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Highlighted Research Publications
 
Citation Abstract Featured Stocks

Beyond punnet squares: student word association and explanations of phenotypic variation through an integrative quantitative genetics unit investigating anthocyanin inheritance and expression in Brassica rapa Fast Plants

2014

Batzli et al.

CBE Life Sci Educ. 13:3:410-424
doi:10.1187/cbe.13-12-0232

Genetics instruction in introductory biology is often confined to Mendelian genetics and avoids the complexities of variation in quantitative traits. Given the driving question “What determines variation in phenotype (Pv)? (Pv=Genotypic variation Gv + environmental variation Ev),” we developed a 4-wk unit for an inquiry-based laboratory course focused on the inheritance and expression of a quantitative trait in varying environments. We utilized Brassica rapa Fast Plants as a model organism to study variation in the phenotype anthocyanin pigment intensity. As an initial curriculum assessment, we used free word association to examine students’ cognitive structures before and after the unit and explanations in students’ final research posters with particular focus on variation (Pv = Gv + Ev). Comparison of pre- and postunit word frequency revealed a shift in words and a pattern of co-occurring concepts indicative of change in cognitive structure, with particular focus on “variation” as a proposed threshold concept and primary goal for students’ explanations. Given review of 53 posters, we found ∼50% of students capable of intermediate to high-level explanations combining both Gv and Ev influence on expression of anthocyanin intensity (Pv). While far from “plug and play,” this conceptually rich, inquiry-based unit holds promise for effective integration of quantitative and Mendelian genetics.

 

DNA-based genetic markers for rapid cycling Brassica rapa (Fast Plants type) designed for the teaching laboratory

2012

Slankster et al.

Front. Plant Sci. 3:118
doi:10.3389/fpls.2012.00118

We have developed DNA-based genetic markers for rapid cycling Brassica rapa (RCBr), also known as Fast Plants. Although markers for B. rapa already exist, ours were intentionally designed for use in a teaching laboratory environment. The qualities we selected for were robust amplification in PCR, polymorphism in RCBr strains, and alleles that can be easily resolved in simple agarose slab gels. We have developed two single nucleotide polymorphism (SNP) based markers and 14 variable number tandem repeat (VNTR)-type markers spread over four chromosomes. The DNA sequences of these markers represent variation in a wide range of genomic features. Among the VNTR-type markers, there are examples of variation in a non-genic region, variation within an intron, and variation in the coding sequence of a gene. Among the SNP-based markers there are examples of polymorphism in intronic DNA and synonymous substitution in a coding sequence. Thus these markers can serve laboratory exercises in both transmission genetics and molecular biology.

 

Mathematical modeling of positron emission tomography (PET) data to assess radiofluoride transport in living plants following petiolar administration

2015

Converse et al.

Plant Methods 11:18
doi:10.1186/s13007-015-0061-y

Ion transport is a fundamental physiological process that can be studied non-invasively in living plants with radiotracer imaging methods. Fluoride is a known phytotoxic pollutant and understanding its transport in plants after leaf absorption is of interest to those in agricultural areas near industrial sources of airborne fluoride. Here we report the novel use of a commercial, high-resolution, animal positron emission tomography (PET) scanner to trace a bolus of [18F]fluoride administered via bisected petioles of Brassica oleracea, an established model species, to simulate whole plant uptake of atmospheric fluoride. This methodology allows for the first time mathematical compartmental modeling of fluoride transport in the living plant. Radiotracer kinetics in the stem were described with a single-parameter free- and trapped-compartment model and mean arrival times at different stem positions were calculated from the free-compartment time-activity curves.

After initiation of administration at the bisected leaf stalk, [18F] radioactivity climbed for approximately 10 minutes followed by rapid washout from the stem and equilibration within leaves. Kinetic modeling of transport in the stem yielded a trapping rate of 1.5 +/− 0.3%/min (mean +/− s.d., n = 3), velocity of 2.2 +/− 1.1 cm/min, and trapping fraction of 0.8 +/− 0.5%/cm.

Quantitative assessment of physiologically meaningful transport parameters of fluoride in living plants is possible using standard positron emission tomography in combination with petiolar radiotracer administration. Movement of free fluoride was observed to be consistent with bulk flow in xylem, namely a rapid and linear change in position with respect to time. Trapping, likely in the apoplast, was observed. Future applications of the methods described here include studies of transport of other ions and molecules of interest in plant physiology.

 

Teaching recurrent selection in the classroom with Wisconsin Fast Plants

1999

Irwin Goldman

HortTechnology 9:4:579-584

Plant breeding is a process that is difficult to compress into laboratory exercises for the classroom. At the heart of plant breeding is the act of selection, a process whereby differential reproduction and survival leads to changes in gene frequency. Given the relatively short span of an academic semester, it has been difficult for students to gain experience with the practice of selection using plant materials. Nearly 15 years ago, P.H. Williams developed Wisconsin Fast Plants, a model system for teaching plant biology in a classroom setting. Wisconsin fast plants are rapid-cycling versions of various Brassica species amenable to a variety of genetic studies due to their short life cycle and ease of handling. This paper describes the development of a model system using Brassica rapa L. fast plants for teaching the cyclical selection process known as recurrent selection in the context of a course on plant breeding. The system allows for up to three cycles of recurrent selection during a single 15-week semester and enables students to gain experience in planting, selection, pollination, and seed harvest during each cycle. With appropriate trait choice, phenotypic changes resulting from selection can be visualized after just three cycles. Using the Fast Plant model, recurrent selection can be practiced successfully by students in the classroom.

 

Rapid-cycling populations of Brassica

1986

Williams et al.

Science 232:4756:1385-1389
doi:10.1126/science.232.4756.1385

Rapid-cycling populations of six economically important species in the genus Brassica have unusual potential for resolving many problems in plant biology and for use in education. Rapid-cycling brassicas can produce up to ten generations of seed per year and serve as models for research in genetics, host-parasite relations, molecular biology, cell biology, plant biochemistry, population biology, and plant breeding. Brassicas are a highly diverse group of crop plants that have great economic value as vegetables and as sources of condiment mustard, edible and industrial oil, animal fodder, and green manure. These plants can also be used in the classroom as convenient, rapidly responding, living plant materials for "hands on" learning at all levels of our educational system.

 

 

Citation Abstract

The gene encoding dihydroflavonol 4- reductase is a candidate for the anthocyaninless locus of rapid cycling Brassica rapa (Fast Plants type)

2016

Wendell et al.

PLoS ONE 11:8
doi:10.1371/journal.pone.0161394

Rapid cycling Brassica rapa, also known as Wisconsin Fast Plants, are a widely used organism in both K-12 and college science education. They are an excellent system for genetics laboratory instruction because it is very easy to conduct genetic crosses with this organism, there are numerous seed stocks with variation in both Mendelian and quantitative traits, they have a short generation time, and there is a wealth of educational materials for instructors using them. Their main deficiency for genetics education is that none of the genetic variation in RCBr has yet been characterized at the molecular level. Here we present the first molecular characterization of a gene responsible for a trait in Fast Plants. The trait under study is purple/nonpurple variation due to the anthocyaninless locus, which is one of the Mendelian traits most frequently used for genetics education with this organism. We present evidence that the DFR gene, which encodes dihyroflavonol 4-reductase, is the candidate gene for the anthocyaninless (ANL) locus in RCBr. DFR shows complete linkage with ANL in genetic crosses with a total of 948 informative chromosomes, and strains with the recessive nonpurple phenotype have a transposon-related insertion in the DFR which is predicted to disrupt gene function. 

On the potential strength and consequences for nonrandom gene flow caused by local adaptation in flowering time

2015

Arthur E Weis

J. Evol. Biol. 28: 699–714. doi:10.1111/jeb.12612

Gene flow is generally considered a random process, that is the loci under consideration have no effect on dispersal success. Edelaar and Bolnick (Trends Ecol Evol, 27, 2012 659) recently argued that nonrandom gene flow could exert a significant evolutionary force. It can, for instance, ameliorate the maladaptive effects of immigration into locally adapted populations. I examined the potential strength for nonrandom gene flow for flowering time genes, a trait frequently found to be locally adapted. The idea is that plants that successfully export pollen into a locally adapted resident population will be a genetically biased subset of their natal population – they will have resident-like flowering times. Reciprocally, recipients will be more migrant-like than the resident population average. I quantified the potential for biased pollen exchange among three populations along a flowering time cline in Brassica rapa from southern California. A two-generation line cross experiment demonstrated genetic variance in flowering time, both within and among populations. Calculations based on the variation in individual flowering schedules showed that resident plants with the most migrant-like flowering times could expect to have up to 10 times more of the their flowers pollinated by immigrant pollen than the least migrant-like. Further, the mean flowering time of the pollen exporters that have access to resident mates differs by up to 4 weeks from the mean in the exporters’ natal population. The data from these three populations suggest that the bias in gene flow for flowering time cuts the impact on the resident population by as much as half. This implies that when selection is divergent between populations, migrants with the highest mating success tend to be resident-like in their flowering times, and so, fewer maladaptive alleles will be introduced into the locally adapting gene pool.

Cotyledon-generated auxin is required for shade-induced hypocotyl growth in Brassica rapa

2014

Procko et al.

Plant Physiology 165:3:1285-1301
doi:10.1104/pp.114.241844

Plant architecture is optimized for the local light environment. In response to foliar shade or neighbor proximity (low red to far-red light), some plant species exhibit shade-avoiding phenotypes, including increased stem and hypocotyl growth, which increases the likelihood of outgrowing competitor plants. If shade persists, early flowering and the reallocation of growth resources to stem elongation ultimately affect the yield of harvestable tissues in crop species. Previous studies have shown that hypocotyl growth in low red to far-red shade is largely dependent on the photoreceptor phytochrome B and the phytohormone auxin. However, where shade is perceived in the plant and how auxin regulates growth spatially are less well understood. Using the oilseed and vegetable crop species Brassica rapa, we show that the perception of low red to far-red shade by the cotyledons triggers hypocotyl cell elongation and auxin target gene expression. Furthermore, we find that following shade perception, elevated auxin levels occur in a basipetal gradient away from the cotyledons and that this is coincident with a gradient of auxin target gene induction. These results show that cotyledon-generated auxin regulates hypocotyl elongation. In addition, we find in mature B. rapa plants that simulated shade does not affect seed oil composition but may affect seed yield. This suggests that in field settings where mutual shading between plants may occur, a balance between plant density and seed yield per plant needs to be achieved for maximum oil yield, while oil composition might remain constant.

Inheritance of rapid cycling in Brassica rapa Fast Plants: dominance that increases with photoperiod

2014

Arthur E Weis

Intl J. Plant Sci. 176:9:859-868
doi:10.1086/683304

Premise of research: The Wisconsin Fast Plants lines of Brassica rapa (RCBr) have been a useful model system for plant ecology, evolution, physiology, and development. However, inheritance of flowering time in the B. rapa Fast Plants has not been explored.

Methodology: I used quantitative genetics to explore additive, dominant, and epistatic genetic components of between-line variance in flowering time and for genotype × photoperiod interaction effects in crosses between RCBr and naturalized Californian populations.

Pivotal results: Strong directional dominance for rapid flowering was evident in the F1 and F2 generations. Evidence was equivocal for epistatic genetic variance between the RCBr and California parental types. The expression of additive genetic variance for flowering time in the naturalized California populations was not masked when combined with the RCBr genetic background. A strong genotype × photoperiod interaction was found; whereas flowering time for RCBr was unaffected by day length, flowering time was halved for California plants under a 24-h photoperiod compared with that under their natural photoperiod regime. Importantly, rapid cycling was completely dominant under constant light but partially dominant under shorter photoperiods.

Conclusions: Early flowering in the commonly used laboratory-derived lines of B. rapa compared with wild populations involves directional dominance that is intensified under long photoperiods. The dominance of accelerated flowering in this case contrasts with the different inheritance patterns seen in independently derived rapid cycling B. rapa lines. Generally, the apparent multiplicity of pathways available for flowering time evolution may contribute to the high incidence of local adaptation in this trait. 

Reaction of lines of the rapid cycling Brassica collection and Arabidopsis thaliana to four pathotypes of Plasmodiophora brassicae

2013

Sharma et al.

Plant Disease 97:6:720-727
doi:10.1094/pdis-08-12-0752-re

The clubroot reaction of five Rapid Cycling Brassica Collection (RCBC) lines (Brassica carinata, B. juncea, B. napus, B. oleracea, and B. rapa) and 84 lines of Arabidopsis thaliana to pathotypes 2, 3, 5, and 6 of Plasmodiophora brassicae (as classified on William’s system) was assessed. Also, the reaction of the Arabidopsis lines to a single-spore isolate of each of pathotypes 3 and 6 was compared with that of a field isolate. Seedlings were inoculated with resting spores of P. brassicae, maintained at 25 and 20°C (day and night, respectively), and assessed for clubroot incidence and severity at 6 weeks after inoculation. Several lines of A. thaliana and RCBC exhibited a differential response to pathotype but none of the lines were immune. Among the RCBC lines, B. napus was resistant to all of the pathotypes; B. oleracea was resistant to pathotypes 2, 3, and 5; B carinata and B. rapa were resistant to pathotypes 2 and 5; and B. juncea was susceptible to pathotypes 5 and 6 and had an intermediate response to pathotypes 2 and 3. Line Ct- 1 of A. thaliana was highly resistant to pathotype 2, Pu2-23 was highly resistant to pathotype 5, and Ws-2 and Sorbo were highly resistant to pathotype 6. These results indicate that the lines of RCBC and A. thaliana have potential for use as model crops for a wide range of studies on clubroot, and could be used to differentiate these four pathotypes of P. brassicae. The reaction of the RCBC lines to pathotype 6 was highly correlated with response under field conditions but the reaction to the single-spore isolates of pathotypes 3 and 6 was not strongly correlated with reaction to the field collections in the Arabidopsis lines.

A comparison of clubroot development and management on canola and Brassica vegetables

2013

Gossen et al.

Canadian J. Plant Path. 35:2:175-191
doi:10.1080/07060661.2013.763293

Clubroot of canola was identified for the first time on the Canadian prairies in 2003, and is spreading rapidly across the region. Although clubroot has been studied extensively on vegetable Brassica crops, it was not clear initially how much of the information would be directly transferable from the intensive production of vegetable crops to the extensive production practices used for canola. This review examines similarities and differences between clubroot development and management on vegetable crops and canola. One important difference was that clubroot generally has a larger economic impact on canola, which is harvested for seed, than on vegetables, especially those where early vegetative growth is the marketable component. Also, clubroot has spread within the production area more quickly than was expected based on vegetable production. This occurs in large part because the resting spores are readily moved within and between fields on the heavy machinery used for canola production, but movement of spores by wind and water is also being assessed. Interestingly, crop rotation to reduce yield losses may be a more viable approach for canola than in vegetable production. Resistance to clubroot is the most consistent and economically viable approach to clubroot management in canola, but several lines of evidence indicate that this resistance may not be durable. Fortunately, the large acreage of canola production in Canada ensures that new sources of resistance will be developed and deployed as existing sources are eroded. Pathogen development and cultural control are very similar on vegetables and canola; bait crops and soil amendments are generally not commercially viable in either system; and biocontrol has a limited potential at this time. Manipulation of seeding date, application of fungicide, and soil fumigation generally have more potential for use in vegetable production than for canola. Identification of approaches that reduce disease pressure in clubroot-infested fields and increase the durability and diversity of genes for clubroot resistance represent important lines of future research.

Clubroot in the Canadian canola crop: 10 years into the outbreak

2013

Strelkov et al.

Canadian J. Plant Path. 36:1:27-36
doi:10.1080/07060661.2013.863807

Clubroot, caused by the obligate parasite Plasmodiophora brassicae, is an important soilborne disease of the Brassicaceae. In Canada, clubroot has long been a problem in cruciferous vegetables, but was not reported on the Prairie canola (Brassica napus) crop until 2003, when 12 clubroot-infested fields were identified in central Alberta. Continued surveillance has shown that the disease is spreading, and at least 1064 fields in the province had confirmed clubroot infestations as of 2012. While the outbreak is still mainly confined to central Alberta, isolated clubroot infestations and the presence of viable P. brassicae inoculum have been confirmed in southern Alberta, Saskatchewan and Manitoba. Dissemination of the parasite appears to be predominantly through the movement of infested soil on farm and other machinery, although secondary mechanisms of spread, such as by wind and water erosion and soil contamination of seeds, also have been implicated. Numerous strains of P. brassicaeoccur in Canada, but pathotype 3 or 16/15/12, as classified on the differentials of Williams or the European Clubroot Differential series, predominates on canola. Given the significant economic value of Canadian canola, the emergence of clubroot on this crop has caused major concern and led to the initiation of a large, coordinated research effort aimed at understanding and managing this disease. This review summarizes the extent and nature of the clubroot outbreak in the Canadian canola crop, 10 years after it began, and provides a context for ongoing research and management efforts.

Reaction to Plasmodiophora brassicae pathotype 6 in lines of Brassica vegetables, Wisconsin Fast Plants, and canola

2012

Adhikari et al.

HortScience 47:3:374-377

Field trials were conducted from 2008 to 2010 to assess the disease reaction to clubroot, caused by Plasmodiophora brassicae Woronin, in selected lines of Brassica spp., including short-season vegetable crops [Shanghai pak choy (B. rapa subsp. Chinensis var. communis)], Chinese flowering cabbage (B. rapa subsp. Chinensis var. utilis), and napa cabbage (B. rapa subsp. Pekinensis), the Rapid Cycling Brassica Collection (RCBC), also known as Wisconsin Fast Plants, and spring canola (B. napus). The trials were conducted on naturally infested soil with P. brassicae at the Muck Crops Research Station in Ontario, Canada, where pathotype 6 is predominant. Clubroot incidence and severity were higher in 2008 and 2010 compared with 2009. The lines of Shanghai pak choy and Chinese flowering cabbage were highly susceptible to clubroot, but each of the clubroot-resistant cultivars of napa cabbage, ‘Deneko’, ‘Bilko’, and ‘Yuki’, was highly resistant to pathotype 6. Among the RCBC lines, B. carinata and B. juncea were highly susceptible and could be used as susceptible models for further studies. Two RCBC lines, B. napus and R. sativus, were resistant to pathotype 6. Two of the canola cultivars, 46A76 and 46A65, were susceptible, but two others, ‘45H21’ and ‘Invigor 5020LL’, were highly resistant to pathotype 6. This difference in response can be exploited in future studies of clubroot reaction in canola. 

Integrating photosynthesis, respiration, biomass partitioning, and plant growth: developing a Microsoft Excel-based simulation model of Wisconsin Fast Plant (Brassica rapa, Brassicaceae) growth with undergraduate students

2011

Grossman et al.

Math. Model. Nat. Phenom. 6:6:295-313
doi:10.1051/mmnp/20116616

This paper demonstrates the development of a simple model of carbon flow during plant growth. The model was developed by six undergraduate students and their instructor as a project in a plant ecophysiology course. The paper describes the structure of the model including the equations that were used to implement it in Excel®, the plant growth experiments that were conducted to obtain information for parameterizing and testing the model, model performance, student responses to the modeling project, and potential uses of the model by other students.

Costs of defense: correlated responses to divergent selection for foliar glucosinolate content in Brassica rapa

2011

Kirk et al.

Evol. Ecol. 25:4:763-775
doi:10.1007/s10682-010-9443-9

The evolutionary response of plant populations to herbivore imposed selection for defense may theoretically be constrained by the costs of defense, yet few studies convincingly demonstrate such costs. We investigated possible constraints on the evolution of defense in rapid cycling Brassica rapa by divergently selecting lines for investment in foliar glucosinolate content, a chemical defense in this species. Costs would then result in a significant correlated response to artificially imposed selection in the direction opposite to the direct response of foliar glucosinolate production. Correlated responses of date of first flowering, total flower number, number of seeds per fruit, and mean seed mass were examined. After three generations of selection, there was a significant direct response in glucosinolate content of the leaves of B. rapa. Furthermore, we found significant correlated responses in both total flower production and number of seeds produced per fruit, but not date of first flowering or mean seed mass. Lines selected for high glucosinolates produced fewer flowers and seeds per fruit compared to those selected for low glucosinolates while lines selected for low glucosinolates showed the opposite response. Thus, costs of defense were demonstrated and may constrain the evolution of foliar glucosinolate production in this plant species.

Uptake of human pharmaceuticals by plants grown under hydroponic conditions

2010

Herklotz et al.

Chemosphere 78:11:1416-1421
doi:10.1016/j.chemosphere.2009.12.048

Cabbage (Brassica rapa var. pekinensis) and Wisconsin Fast Plants (Brassica rapa) were chosen for a proof of concept study to determine the potential uptake and accumulation of human pharmaceuticals by plants. These plants were grown hydroponically under high-pressure sodium lamps in one of two groups including a control and test group exposed to pharmaceuticals. The control plants were irrigated with a recirculating Hoagland’s nutrient solution while the test plants were irrigated with a Hoagland’s nutrient solution fortified with the pharmaceuticals carbamazepine, salbutamol, sulfamethoxazole, and trimethoprim at 232.5 lg L1 . When plants reached maturity, nine entire plants of each species were separated into components such as roots, leafs, stems, and seedpods where applicable. An analytical method for quantifying pharmaceuticals and personal care products was developed using pressurized liquid extraction and liquid chromatography electrospray ionization mass spectrometry (LC/ESI/MS) in positive and negative ion modes using single ion monitoring. The method detection limits ranged from 3.13 ng g1 to 29.78 ng g1 with recoveries ranging from 66.83% to 113.62% from plant matrices. All four of the pharmaceuticals were detected in the roots and leaves of the cabbage. The maximum wet weight concentrations of the pharmaceuticals were detected in the root structure of the cabbage plants at 98.87 ng g1 carbamazepine, 114.72 ng g1 salbutamol, 138.26 ng g1 sulfamethoxazole, and 91.33 ng g1 trimethoprim. Carbamazepine and salbutamol were detected in the seedpods of the Wisconsin Fast Plants while all four of the pharmaceuticals were detected in the leaf/stem/root of the Wisconsin Fast Plants. Phloroglucinol staining of root cross-sections was used to verify the existence of an intact endodermis, suggesting that pharmaceuticals found in the leaf and seedpods of the plants were transported symplastically.

Adventitious shoot regeneration from cotyledonary explants of rapid cycling Fast Plants of Brassica rapa L.

2010

Cogbill et al.

Plant Cell Tissue Organ Cult. 101:2:127-133
doi:10.1007/s11240-010-9669-9

Rapid-cycling fast plants (Brassica rapa; RCBr) is also known as Wisconsin Fast Plant and is widely used in K-12 and undergraduate studies. RCBr has a short generation time (seed-to-seed in 30–60 days), which allows for the completion of experiments in a semester. Previous studies have shown that cotyledonary explants with attached petioles are capable of generating shoots. However, there is no published adventitious shoot regeneration protocol to date. Sterile cotyledonary explants were excised; all edges and petioles were removed. Five-day-old cotyledonary explants produced shoots on a Murashige and Skoog medium containing 1.5 mg/L thiadiazuron (TDZ) and 0.5 mg/L 1-naphthaleneacetic acid (NAA) (FPM I) at a mean rate of 8.8%. This rate increased to 14.8% in explants placed on FPM I medium supplemented with 5.0 mg/L silver nitrate (AgNO3) (SRM 2). The rate increased to 32.5% when 5-day-old explants, excised from the part of the cotyledon nearest to the petiole, were placed adaxial side up on SRM 2 medium. The shoot regeneration rate increased to 44.5% using 4-day-old cotyledonary explants. A shoot regeneration rate of 23% was observed among 9-day-old leaf explants. Shoots from cotyledonary explants were elongated on basal medium with 0.5 mg/L NAA, rooted on basal medium, and later acclimatized. This is the first report of shoot regeneration from cotyledonary explants of rapid-cycling Brassica rapa without pre-existing meristematic tissues.

Development of public immortal mapping populations, molecular markers and linkage maps for rapid cycling Brassica rapa and B. oleracea

2009

Iniguez-Luy et al.

Theor. Appl. Genet. 120:1:31-43
doi:10.1007/s00122-009-1157-4

Publicly available genomic tools help researchers integrate information and make new discoveries. In this paper, we describe the development of immortal mapping populations of rapid cycling, self-compatible lines, molecular markers, and linkage maps for Brassica rapa and B. oleracea and make the data and germplasm available to the Brassica research community. The B. rapa population consists of 160 recombinant inbred (RI) lines derived from the cross of highly inbred lines of rapid cycling and yellow sarson B. rapa. The B. oleracea population consists of 155 double haploid (DH) lines derived from an F1 cross between two DH lines, rapid cycling and broccoli. A total of 120 RFLP probes, 146 SSR markers, and one phenotypic trait (flower color) were used to construct genetic linkage maps for both species. The B. rapa map consists of 224 molecular markers distributed along 10 linkage groups (A1–A10) with a total distance of 1125.3 cM and a marker density of 5.7 cM/marker. The B. oleracea genetic map consists of 279 molecular markers and one phenotypic marker distributed along nine linkage groups (C1–C9) with a total distance of 891.4 cM and a marker density of 3.2 cM/marker. A syntenic analysis with Arabidopsis thaliana identified collinear genomic blocks that are in agreement with previous studies, reinforcing the idea of conserved chromosomal regions across the Brassicaceae.

Forward and reverse genetics of rapid cycling Brassica oleracea

2009

Himelblau et al.

Theor. Appl. Genet. 118:5:953-961
doi:10.1007/s00122-008-0952-7

Seeds of rapid-cycling Brassica oleracea were mutagenized with the chemical mutagen, ethylmethane sulfonate. The reverse genetics technique, TILLING, was used on a sample population of 1,000 plants, to determine the mutation profile. The spectrum and frequency of mutations induced by ethylmethane sulfonate was similar to that seen in other diploid species such as Arabidopsis thaliana. These data indicate that the mutagenesis was effective and demonstrate that TILLING represents an efficient reverse genetic technique in B. oleracea that will become more valuable as increasing genomic sequence data become available for this species. The extensive duplication in the B. oleracea genome is believed to result in the genetic redundancy that has been important for the evolution of morphological diversity seen in today’s B. oleracea crops (broccoli, Brussels sprouts, cauliflower, cabbage, kale and kohlrabi). However, our forward genetic screens identified 120 mutants in which some aspect of development was affected. Some of these lines have been characterized genetically and in the majority of these, the mutant trait segregates as a recessive allele affecting a single locus. One dominant mutation (curly leaves) and one semi-dominant mutation (dwarf-like) were also identified. Allelism tests of two groups of mutants (glossy and dwarf) revealed that for some loci, multiple independent alleles have been identified. These data indicate that, despite genetic redundancy, mutation of many individual loci in B. oleracea results in distinct phenotypes.

Development of a set of public SSR markers derived from genomic sequence of a rapid cycling Brassica oleracea L. genotype

2008

Iniguez-Luy et al.

Theor. Appl. Genet. 117:6:977-985
doi:10.1007/s00122-008-0837-9

The traditional development of simple sequence repeat (SSR) or microsatellite markers by probe hybridization can be time-consuming and requires the use of specialized laboratory equipment. In this study, probe hybridization was circumvented by using sequence information on 3,500 genomic clones mainly from Brassica oleracea to identify di, tri, tetra and penta-nucleotide repeats. A total of 587 primer pairs flanking SSR were developed using this approach. From these, 420 SSR markers amplified DNA in two parental lines of B. rapa (26% were polymorphic) and 523 in two parental lines of B. oleracea (32% were polymorphic). A diverse array of motif types was identified, characterized and compared with traditional SSR detection methods. The most abundant motifs found were di- (38%) and trinucleotides (33%) followed by penta- (16%) and tetranucleotide (13%) motifs. The type of motif class, motif length and repeat were not indicative of polymorphisms. The frequency of B. oleracea SSRs in genomic shotgun sequence was estimated to be 1 every 4 Kb. In general, the average motif length and repeat numbers were shorter than those obtained previously by probe hybridization, and they contained a more balanced representation of SSR motif types in the genome by identifying those that do not hybridize well to DNA probes. Brassica genomic DNA sequence information is a promising resource for developing a large number of SSR molecular markers in Brassica species.

Effects of stress and phenotypic variation on inbreeding depression in Brassica rapa

2008

Waller et al.

Evolution 62:4:917-931
doi:10.1111/j.1558-5646.2008.00325.x

Stressful environments are often said to increase the expression of inbreeding depression. Alternatively, Crow's “opportunity for selection” (the squared phenotypic coefficient of variation) sets a limit to how much selection can occur, constraining the magnitude of inbreeding depression. To test these hypotheses, we planted self- and cross-fertilized seeds of Brassica rapa into a factorial experiment that varied plant density and saline watering stresses. We then repeated the experiment, reducing the salt concentration. We observed considerable inbreeding depression, particularly for survival in the first experiment and growth in the second. Both stresses independently depressed plant performance. Families differed in their amounts of inbreeding depression and reaction norms across environments. Outcrossed progeny were sometimes more variable. Stresses had small and inconsistent effects on inbreeding depression and, when significant, tended to diminish it. Levels of phenotypic variability often predicted whether inbreeding depression would increase or decrease across environments and were particularly effective in predicting which traits display the most inbreeding depression. Thus, we find little support for the stress hypothesis and mixed support for the phenotypic variability hypothesis. Variable levels of phenotypic variation provide a parsimonious explanation for shifts in inbreeding depression that should be tested before invoking more complex hypotheses.

Teaching human genetics with mustard: rapid cycling Brassica rapa (Fast Plants type) as a model for human genetics in the classroom laboratory

2007

Wendell et al.

CBE Life Sci. Educ. 6:2:179-185
doi:10.1187/cbe.07-02-0010

We have developed experiments and materials to model human genetics using rapid cycling Brassica rapa, also known as Fast Plants. Because of their self-incompatibility for pollination and the genetic diversity within strains, B. rapa can serve as a relevant model for human genetics in teaching laboratory experiments. The experiment presented here is a paternity exclusion project in which a child is born with a known mother but two possible alleged fathers. Students use DNA markers (microsatellites) to perform paternity exclusion on these subjects. Realistic DNA marker analysis can be challenging to implement within the limitations of an instructional lab, but we have optimized the experimental methods to work in a teaching lab environment and to maximize the “hands-on” experience for the students. The genetic individuality of each B. rapa plant, revealed by analysis of polymorphic microsatellite markers, means that each time students perform this project, they obtain unique results that foster independent thinking in the process of data interpretation.

Ozone affects gas exchange, growth, and reproductive development in Brassica camprestris (Wisconsin Fast Plants)

2007

Black et al.

New Phytologist 176:1:150-163
doi:10.1111/j.1469-8137.2007.02163.x

Exposure to ozone (O3) may affect vegetative and reproductive development, although the consequences for yield depend on the effectiveness of the compensatory processes induced. This study examined the impact on reproductive development of exposing Brassica campestris (Wisconsin Fast Plants) to ozone during vegetative growth.

Plants were exposed to 70 ppb ozone for 2 d during late vegetative growth or 10 d spanning most of the vegetative phase. Effects on gas exchange, vegetative growth, reproductive development and seed yield were determined.

Impacts on gas exchange and foliar injury were related to pre-exposure stomatal conductance. Exposure for 2 d had no effect on growth or reproductive characteristics, whereas 10-d exposure reduced vegetative growth and reproductive site number on the terminal raceme. Mature seed number and weight per pod and per plant were unaffected because seed abortion was reduced.

The observation that mature seed yield per plant was unaffected by exposure during the vegetative phase, despite adverse effects on physiological, vegetative and reproductive processes, shows that indeterminate species such as B. campestris possess sufficient compensatory flexibility to avoid reductions in seed production.

Rapid evolution of flowering time by an annual plant in response to a climate fluctuation

2007

Franks et al.

PNAS 104:4:1278-1282
doi:10.1073/pnas.0608379104

Ongoing climate change has affected the ecological dynamics of many species and is expected to impose natural selection on ecologically important traits. Droughts and other anticipated changes in precipitation may be particularly potent selective factors, especially in arid regions. Here we demonstrate the evolutionary response of an annual plant, Brassica rapa, to a recent climate fluctuation resulting in a multiyear drought. Ancestral (predrought) genotypes were recovered from stored seed and raised under a set of common environments with descendant (postdrought) genotypes and with ancestor×descendant hybrids. As predicted, the abbreviated growing seasons caused by drought led to the evolution of earlier onset of flowering. Descendants bloomed earlier than ancestors, advancing first flowering by 1.9 days in one study population and 8.6 days in another. The intermediate flowering time of ancestor×descendant hybrids supports an additive genetic basis for divergence. Experiments confirmed that summer drought selected for early flowering, that flowering time was heritable, and that selection intensities in the field were more than sufficient to account for the observed evolutionary change. Natural selection for drought escape thus appears to have caused adaptive evolution in just a few generations. A systematic effort to collect and store propagules from suitable species would provide biologists with materials to detect and elucidate the genetic basis of further evolutionary shifts driven by climate change.

Mapping the Anthocyaninless (anl) locus in rapid cycling Brassica rapa (RBr) to linkage group R9

2007

Burdzinski et al.

BMC Genetics 8:64
doi:10.1186/1471-2156-8-64

Anthocyanins are flavonoid pigments that are responsible for purple coloration in the stems and leaves of a variety of plant species. Anthocyaninless (anl) mutants of Brassica rapa fail to produce anthocyanin pigments. In rapid-cycling Brassica rapa, also known as Wisconsin Fast Plants, the anthocyaninless trait, also called non-purple stem, is widely used as a model recessive trait for teaching genetics. Although anthocyanin genes have been mapped in other plants such as Arabidopsis thaliana, the anl locus has not been mapped in any Brassica species.

We tested primer pairs known to amplify microsatellites in Brassicas and identified 37 that amplified a product in rapid-cycling Brassica rapa. We then developed three-generation pedigrees to assess linkage between the microsatellite markers and anl. 22 of the markers that we tested were polymorphic in our crosses. Based on 177 F2 offspring, we identified three markers linked to anl with LOD scores ≥ 5.0, forming a linkage group spanning 46.9 cM. Because one of these markers has been assigned to a known B. rapa linkage group, we can now assign the anl locus to B. rapa linkage group R9.

This study is the first to identify the chromosomal location of an anthocyanin pigment gene among the Brassicas. It also connects a classical mutant frequently used in genetics education with molecular markers and a known chromosomal location.

Effects of nutrient and CO2 availability on tolerance to herbivory in Brassica rapa 

2007

Marshall et al.

Plant Ecology 196:1:1-13
doi:10.1007/s11258-007-9331-8 

The ability of plants to recover from herbivore damage and maintain their fitness depends on physiological mechanisms that are affected by the availability of resources such as carbon and soil nutrients. In this study, we explored the effects of increased carbon and nutrient availability on the response of rapid cycling Brassica rapa to damage by the generalist herbivore, Trichoplusia ni (Noctuidae), in a greenhouse experiment. Using fruit mass as an estimate of plant fitness, we tested three physiological models, which predict either an increase or a decrease of tolerance to herbivory with increasing resource availability. We used leaf demography to examine some plausible mechanisms through which resource availability may affect tolerance. Our results contradict all models, and, rather, they support a more complicated view of the plasticity of resource uptake and allocation than the ones considered by the models tested. Fruit mass was negatively affected by herbivore damage only under elevated CO2, and only for certain harvest dates. Increased CO2 had no effect on the number of leaf births, but it decreased leaf longevity and the total number of leaves on a plant. Nutrient addition increased the number of leaf births, leaf longevity and the total number of leaves on a plant. We conclude that a shortening of the life span of the plants, brought about by elevated CO2, was responsible for a higher susceptibility of plants to herbivore damage under high CO2 concentration.

Selenium increases sulfur uptake and regulates glucosinolate metabolism in rapid cycling Brassica oleracea

2007

Toler et al.

JASHS 132:1:14-19

Glucosinolates are sulfur-containing secondary plant metabolites commonly found in the family Brassicaceae. The presence of selenium in soils increases the uptake of sulfur and inhibits the production of glucosinolates in brassicaceous plants. This study was undertaken to determine the extent of selenium's impact on sulfur uptake and glucosinolate production in Brassica oleracea L. Rapid-cycling B. oleracea plants were grown hydroponically in half-strength Hoagland's nutrient solution with selenium treatments delivered as sodium selenate concentrations of 0.0, 0.5, 0.75, 1.0, and 1.5 mg·L−1. Elevated sulfur treatments of 37 mg·L−1 sulfate and 37 mg·L−1 sulfate/0.75 mg·L−1 selenate were incorporated to compare with selenium treatments. Plants were harvested and freeze-dried 1 day before anthesis. Selenium and sulfur content of plant tissue was determined by flame atomic absorption spectrophotometry and a carbon–nitrogen–sulfur analyzer. Glucosinolate content of leaf tissue was determined by high-performance liquid chromatography. Selenium and sulfur uptake in plants positively correlated with selenium concentration in the nutrient solution. The sulfur concentration of plants exposed to selenium equaled or exceeded the sulfur concentration of plants exposed to elevated sulfur. Despite higher sulfur concentrations, there occurred a statistically significant decrease in production of five of the seven glucosinolates analyzed in selenium-enriched plants. Plants that underwent elevated sulfur treatments had higher glucosinolate production than selenium-treated plants. These results suggest that selenium either upregulates or prevents the downregulation of sulfur uptake in B. oleracea. In addition, the presence of selenium within the plant appears to have a negative impact on the production of certain glucosinolates despite adequate availability of sulfur.

Characterizing genotype specific differences in survival, growth, and reproduction for field grown, rapid cycling Brassica rapa

2006

Martin G Kelly

Env. Exp. Botany 55:1-2:61-69
doi:10.1016/j.envexpbot.2004.09.012 

Rapid cycling Brassica rapa (RCBr) develops rapidly, and has both small adult size and a brief life cycle. Yet, in spite of many investigations using RCBr, extremely few plant ecologists have used this plant in the field. This study is the first to describe the genotype specific variation in traits describing survival, growth, and reproduction for field grown, RCBr. I also identify traits associated with fitness. Five genotypes of RCBr were used: standard, anthocyaninless, yellow-green, anthocyaninless and hairless, and anthocyaninless and yellow-green. Plants were grown outside in a “common garden”. Eight plant traits were measured: life span, height, growth rate, leaf size, number of flowers and fruits, fruit set, and fitness. All traits, except life span, differed significantly among the five plant genotypes. Correlation analysis revealed that fitness increased as each of these of seven plant traits increased. This study demonstrates that RCBr can serve as a model organism in ecological field studies.

Genetic variation and selection response in model breeding populations of Brassica rapa following a diversity bottleneck

2006

Briggs et al.

Genetics 172:1:457-465
doi:10.1534/genetics.105.040899 

Domestication and breeding share a common feature of population bottlenecks followed by significant genetic gain. To date, no crop models for investigating the evolution of genetic variance, selection response, and population diversity following bottlenecks have been developed. We developed a model artificial selection system in the laboratory using rapid-cycling Brassica rapa. Responses to 10 cycles of recurrent selection for cotyledon size were compared across a broad population founded with 200 individuals, three bottleneck populations initiated with two individuals each, and unselected controls. Additive genetic variance and heritability were significantly larger in the bottleneck populations prior to selection and this corresponded to a heightened response of bottleneck populations during the first three cycles. However, the overall response was ultimately greater and more sustained in the broad population. AFLP marker analyses revealed the pattern and extent of population subdivision were unaffected by a bottleneck even though the diversity retained in a selection population was significantly limited. Rapid gain in genetically more uniform bottlenecked populations, particularly in the short term, may offer an explanation for why domesticators and breeders have realized significant selection progress over relatively short time periods.

Mapping and characterization of FLC homologs and QTL analysis of flowering time in Brassica oleracea

2006

Okazaki et al.

Theor. Appl. Genet. 114:4:595-608
doi:10.1007/s00122-006-0460-6 

The FLC gene product is an inhibitor of flowering in ArabidopsisFLC homologs in Brassica species are thought to control vernalization. We cloned four FLC homologs (BoFLCs) from Brassica oleracea. Three of these, BoFLC1BoFLC3 and BoFLC5, have been previously characterized. The fourth novel sequence displayed 98% sequence homology to the previously identified gene BoFLC4, but also showed 91% homology to BrFLC2 from Brassica rapa. Phylogenetic analysis showed that this clone belongs to the FLC2 clade. Therefore, we designated this gene BoFLC2. Based on the segregation of RFLP, SRAP, CAPS, SSR and AFLP loci, a detailed linkage map of B. oleracea was constructed in the F2 progeny obtained from a cross of B. oleracea cv. Green Comet (broccoli; non-vernalization type) and B. oleracea cv. Reiho (cabbage; vernalization type), which covered 540 cM, 9 major linkage groups. Six quantitative trait loci (QTL) controlling flowering time were detected. BoFLC1BoFLC3 and BoFLC5 were not linked to the QTLs controlling flowering time. However, the largest QTL effect was located in the region where BoFLC2 was mapped. Genotyping of Fplants at the BoFLC2 locus showed that most of the early flowering plants were homozygotes of BoFLC-GC, whereas most of the late- and non-flowering plants were homozygotes of BoFLC-Rei. The BoFLC2 homologs present in plants of the non-vernalization type were non-functional, due to a frameshift in exon 4. Moreover, duplications and deletions of BoFLC2 were detected in broccoli and a rapid cycling line, respectively. These results suggest that BoFLC2 contributes to the control of flowering time in B. oleracea.

Seed coat patterns in rapid cycling Brassica forms

2005

Karcz et al.

ABCSB 47:1:159-165

The micromorphological typology of seed surfaces was investigated in rapid-cycling Brassica (RCBr) forms, using scanning electron microscopy. Four types of basic ornamentation pattern were recognized: reticulate (B. rapa, B. juncea), reticulate-foveolate (B. nigra), randomly reticulate (B. oleracea, B. napus) and reticulate-rugose (B. carinata). The seed coats showed variation in the shape and size of the testa epidermal cells and the structure of the outer periclinal and anticlinal cell walls. The surface patterns in RCBr seeds were less exposed and were moderately reticulated compared to cultivated Brassica species. The micromorphological characteristics of the seed coats may provide valuable additional diagnostic criteria for delimitation of RCBr forms, and can be used in identification of seeds. 

A novel dwarfing mechanism in a green revolution gene from Brassica rapa

2005

Muangprom et al.

Plant Physiology 137:3:931-938
doi:10.1104/pp.104.057646 

Mutations in the biosynthesis or signaling pathways of gibberellin (GA) can cause dwarfing phenotypes in plants, and the use of such mutations in plant breeding was a major factor in the success of the Green Revolution. DELLA proteins are GA signaling repressors whose functions are conserved in different plant species. Recent studies show that GA promotes stem growth by causing degradation of DELLA proteins via the ubiquitin-proteasome pathway. The most widely utilized dwarfing alleles in wheat (Triticum aestivum; e.g. Rht-B1b and Rht-D1b) encode GA-resistant forms of a DELLA protein that function as dominant and constitutively active repressors of stem growth. All of the previously identified dominant DELLA repressors from several plant species contain N-terminal mutations. Here we report on a novel dwarf mutant from Brassica rapa (Brrga1-d) that is caused by substitution of a conserved amino acid in the C-terminal domain of a DELLA protein. Brrga1-d, like N-terminal DELLA mutants, retains its repressor function and accumulates to high levels, even in the presence of GA. However, unlike wild-type and N-terminal DELLA mutants, Brrga1-d does not interact with a protein component required for degradation, suggesting that the mutated amino acid causes dwarfism by preventing an interaction needed for its degradation. This novel mutation confers nondeleterious dwarf phenotypes when transferred to Arabidopsis (Arabidopsis thaliana) and oilseed rape (Brassica napus), indicating its potential usefulness in other crop species.

Seed storage reserves and glucosinolates in Brassica rapa L. grown on the international space station

2005

Musgrave et al.

JASHS 130:6:848-856

Although plants are envisioned to play a central role in life support systems for future long-duration space travel, plant growth in space has been problematic due to horticultural problems of nutrient delivery and gas resupply posed by the weightless environment. Iterative improvement in hardware designed for growth of plants on orbital platforms now provides confidence that plants can perform well in microgravity, enabling investigation of their nutritional characteristics. Plants of B. rapa (cv. Astroplants) were grown in the Biomass Production System on the International Space Station. Flowers were hand-pollinated and seeds were produced prior to harvest at 39 days after planting. The material was frozen or fixed while on orbit and subsequently analyzed in our laboratories. Gross measures of growth, leaf chlorophyll, starch and soluble carbohydrates confirmed comparable performance by the plants in spaceflight and ground control treatments. Analysis of glucosinolate production in the plant stems indicated that 3-butenylglucosinolate concentration was on average 75% greater in flight samples than in ground control samples. Similarly, the biochemical make-up of immature seeds produced during spaceflight and fixed or frozen while in orbit was significantly different from the ground controls. The immature seeds from the spaceflight treatment had higher concentrations of chlorophyll, starch, and soluble carbohydrates than the ground controls. Seed protein was significantly lower in the spaceflight material. Microscopy of immature seeds fixed in flight showed embryos to be at a range of developmental stages, while the ground control embryos had all reached the premature stage of development. Storage reserve deposition was more advanced in the ground control seeds. The spaceflight environment thus influences B. rapa metabolite production in ways that may affect flavor and nutritional quality of potential space produce. 

A method to teach age specific demography with field grown rapid cycling Brassica rapa (Wisconsin Fast Plants)

2004

Kelly et al.

JNRLSE 33:40-46

In this paper, we demonstrate that rapid cycling Brassica rapa (Wisconsin Fast Plants) can be used in inquiry-based, student ecological fieldwork. We are the first to describe age-specific survival for field-grown Fast Plants and identify life history traits associated with individual survival. This experiment can be adapted by educators as a method to teach age-specific demography or ecology with Fast Plants. Four genotypes of Fast Plants (dwarf, elongated internode, standard, and rosette) were used. Fast Plants were grown in a "common garden" experiment. The site was surveyed every day to monitor emergence, and every 2 days to record mortality. Seven life-history traits were measured: days to emergence, emergence date, death date, life span, flowering date, juvenile days, and adult days. Most seedlings emerged 5 to 6 days after sowing and plants lived an average of 30 days. Flowering began about 17 days after seedling emergence. Life table analysis showed that Fast Plants have Type I age-specific survivorship. Analysis of variance showed that these four genotypes did not differ for any of the life-history traits measured. However, correlation analysis revealed that life span was negatively related to emergence date, and flowering date was positively related with emergence date. We suggest that ecologists extend the use of Fast Plants to include student field-work. Through inquiry based work, students can construct a personal understanding about the process, practice, and outcomes of science. Students will also gain understanding about the limits of numerical data, their analysis, and their interpretation. 

Linkage mapping of genes controlling resistance to white rust (Albugo candida) in Brassica rapa (syn. campestris) and comparitive mapping to Brassica napus and Arabidopsis thaliana

2002

Kole et al.

Genome 45:1:22-27
doi:10.1139/g01-123

Genes for resistance to white rust (Albugo candida) in oilseed Brassica rapa were mapped using a recombinant inbred (RI) population and a genetic linkage map consisting of 144 restriction fragment length polymorphism (RFLP) markers and 3 phenotypic markers. Young seedlings were evaluated by inoculating cotyledons with A. candida race 2 (AC2) and race 7 (AC7) and scoring the interaction phenotype (IP) on a 0–9 scale. The IP of each line was nearly identical for the two races and the population showed bimodal distributions, suggesting that a single major gene (or tightly linked genes) controlled resistance to the two races. The IP scores were converted to categorical resistant and susceptible scores, and these data were used to map a single Mendelian gene controlling resistance to both races on linkage group 4 where resistance to race 2 had been mapped previously. A quantitative trait loci (QTL) mapping approach using the IP scores detected the same major resistance locus for both races, plus a second minor QTL effect for AC2 on linkage group 2. These results indicate that either a dominant allele at a single locus (Aca1) or two tightly linked loci control seedling resistance to both races of white rust in the biennial turnip rape cultivar Per. The map positions of white rust resistance genes in B. rapa and Brassica napus were compared and the results indicate where additional loci that have not been mapped may be located. Alignment of these maps to the physical map of the Arabidopsis genome identified regions to target for comparative fine mapping using this model organism. 

Direct and indirect responses to selection on pollen size in Brassica rapa L.

2001

Sarkissian et al.

J. Evol. Botany 14:3:456-468
doi:10.1046/j.1420-9101.2001.00285.x

Pollen size varies little within angiosperm species, but differs extensively between species, suggesting the action of strong selection. Nevertheless, the potential for genetic responses of pollen size to selection, as determined by additive genetic variance and genetic correlations with other floral traits, has received little attention. To assess this potential, we subjected Brassica rapato artificial selection for large and small pollen during three generations. This selection caused significant divergence in pollen diameter, with additive genetic effects accounting for over 30% of the observed phenotypic variation in pollen size. Such heritable genetic variation suggests that natural selection could effect evolutionary change in this trait. Selection on pollen size also elicited correlated responses in pollen number (–), flower size (+), style length (+), and ovule number (+), suggesting that pollen size cannot evolve independently. The correlated responses of pollen number, flower size and ovule number probably reflect the genetically determined and physically constrained pattern of resource allocation in B. rapa. In contrast, the positive correlation between pollen size and style length may represent a widespread gametic-phase disequilibrium in angiosperms that arises from nonrandom fertilization success of large pollen in pistils with long styles.

Molybdenum sequestration in Brassica species, a role for anthocyanins

2001

Hale et al.

Plant Physiology 126:4:1391-1402
doi:10.1104/pp.126.4.1391

To elucidate plant mechanisms involved in molybdenum (Mo) sequestration and tolerance, Brassica spp. seedlings were supplied with molybdate, and the effects on plant physiology, morphology, and biochemistry were analyzed. When supplied with (colorless) molybdate Indian mustard (Brassica juncea) seedlings accumulated water-soluble blue crystals in their peripheral cell layers. Energy dispersive x-ray analysis showed that Mo accumulated predominantly in the vacuoles of the epidermal cells. Therefore, the blue crystals are likely to be a Mo compound. The x-ray absorption spectrum of the plant-accumulated Mo was different than that for molybdate, indicating complexation with a plant molecule. Because the blue compound was water soluble and showed a pH-dependent color change, possible involvement of anthocyanins was investigated. An anthocyanin-less mutant of Brassica rapa (“fast plants”) was compared with varieties containing normal or high anthocyanin levels. The anthocyanin-less mutant did not show accumulation of a blue compound when supplied with molybdate. In the anthocyanin-containing varieties, the blue compound colocalized with anthocyanins in the peripheral cell layers. Mo accumulation by the three B. rapa varieties was positively correlated with anthocyanin content. Addition of molybdate to purified B. rapa anthocyanin resulted in an in vitro color change from pink to blue. Therefore, Mo appears to be sequestered in vacuoles of the peripheral cell layers of Brassica spp. as a blue compound, probably a Mo-anthocyanin complex.

Realizing the potential of rapid cycling Brassica as a model system for use in plant biology research

2000

Mary E Musgrave

J. Plant Growth Regul. 19:3:314-325
doi:10.1007/s003440000036

Rapid-cycling Brassica populations were initially developed as a model for probing the genetic basis of plant disease. Paul Williams and co-workers selected accessions of the six main species for short time to flower and rapid seed maturation. Over multiple generations of breeding and selection, rapid-cycling populations of each of the six species were developed. Because of their close relationship with economically important Brassica species, rapid-cycling Brassica populations, especially those of B. rapa (RCBr) and B. oleracea, have seen wide application in plant and crop physiology investigations. Adding to the popularity of these small, short-lived plants for research applications is their extensive use in K–12 education and outreach.

Gravity independence of seed-to-seed cycling in Brassica rapa

2000

Musgrave et al.

Planta 210:3:400-406
doi:10.1007/PL00008148

Growth of higher plants in the microgravity environment of orbital platforms has been problematic. Plants typically developed more slowly in space and often failed at the reproductive phase. Short-duration experiments on the Space Shuttle showed that early stages in the reproductive process could occur normally in microgravity, so we sought a long-duration opportunity to test gravity's role throughout the complete life cycle. During a 122-d opportunity on the Mir space station, full life cycles were completed in microgravity with Brassica rapa L. in a series of three experiments in the Svet greenhouse. Plant material was preserved in space by chemical fixation, freezing, and drying, and then compared to material preserved in the same way during a high-fidelity ground control. At sampling times 13 d after planting, plants on Mir were the same size and had the same number of flower buds as ground control plants. Following hand-pollination of the flowers by the astronaut, siliques formed. In microgravity, siliques ripened basipetally and contained smaller seeds with less than 20% of the cotyledon cells found in the seeds harvested from the ground control. Cytochemical localization of storage reserves in the mature embryos showed that starch was retained in the spaceflight material, whereas protein and lipid were the primary storage reserves in the ground control seeds. While these successful seed-to-seed cycles show that gravity is not absolutely required for any step in the plant life cycle, seed quality in Brassica is compromised by development in microgravity.

Pollination and embryo development in Brassica rapa L. in microgravity

2000

Kuang et al.

Intl. J Plant Sci. 161:2:203-211
doi:10.1086/314254

Plant reproduction under spaceflight conditions has been problematic in the past. In order to determine what aspect of reproductive development is affected by microgravity, we studied pollination and embryo development in Brassica rapa L. during 16 d in microgravity on the space shuttle (STS‐87). Brassica is self‐incompatible and requires mechanical transfer of pollen. Short‐duration access to microgravity during parabolic flights on the KC‐135A aircraft was used initially to confirm that equal numbers of pollen grains could be collected and transferred in the absence of gravity. Brassica was grown in the Plant Growth Facility flight hardware as follows. Three chambers each contained six plants that were 13 d old at launch. As these plants flowered, thin colored tape was used to indicate the date of hand pollination, resulting in silique populations aged 8–15 d postpollination at the end of the 16‐d mission. The remaining three chambers contained dry seeds that germinated on orbit to produce 14‐d‐old plants just beginning to flower at the time of landing. Pollen produced by these plants had comparable viability (93%) with that produced in the 2‐d‐delayed ground control. Matched‐age siliques yielded embryos of equivalent developmental stage in the spaceflight and ground control treatments. Carbohydrate and protein storage reserves in the embryos, assessed by cytochemical localization, were also comparable. In the spaceflight material, growth and development by embryos rescued from siliques 15 d after pollination lagged behind the ground controls by 12 d; however, in the subsequent generation, no differences between the two treatments were found. The results demonstrate that while no stage of reproductive development in Brassica is absolutely dependent upon gravity, lower embryo quality may result following development in microgravity.