Genetic diversity in a germplasm collection of Brassica rapa subsp rapa L. from northwestern Spain

In Galicia (northwestern Spain), Brassica rapa subsp. rapa L. includes turnips, turnip greens, and turnip tops. The objectives of this study are (i) to determine the genetic diversity and morphological resemblances among the B. rapa landraces of northwestern Spain in order to have information relative to the resources preserved, and (ii) to evaluate their agronomic characteristics, considering the three potential products that can be harvested. A collection of 120 landraces was evaluated for 34 morphological and agronomical traits by an augmented design at two locations. Two landraces were the most promising for turnips production (MBG-BRS0183 and MBG-BRS0256), two showed the best characteristics for turnip greens (MBG-BRS0082 and MBG-BRS0184), and three were the most appropriate landraces for turnip tops production (MBG-BRS0143, MBG-BRS0173 and MBG-BRS0401). Landraces were classified into five clusters (A–E) using the Ward–MLM (Modified Location Model) strategy: (A) included plants with the worst agronomic potential, (B) included most of the turnip populations with rosette growth habit, (C) included turnip populations without rosette growth habit, having more vigorous plants than cluster B, (D) gathered the most vigorous local varieties, with the highest early vigor, fresh matter content per leaf, and number of secondary stems per plant, and (E) landraces characterized by their earliness, large flowering periods, high numbers of secondary stems, and large seed weights. As conclusion, landraces evaluated in this work displayed enough variability to differentiate among appropriate populations for each one of the distinct crops. Their classification, using the Ward–MLM strategy, grouped accessions with similar characteristics into homogeneous categories.     

青海大黄油菜Brsc1基因的精细定位及图谱整合

大黄油菜是源于青海湟源的地方品种,种皮颜色鲜黄,其大黄油菜的黄籽性状受1对隐性基因(Brsc1)控制,该基因被定位于白菜A9染色体上一段1.7 Mb的区间内。为了更好地利用这一黄籽资源,对Brsc1基因进一步精细定位。利用青海大黄油菜和褐籽白菜型油菜09A-126构建BC4及F2分离群体。利用白菜同源区段内已公布的SSR标记,同时利用该区段序列信息开发新的SSR引物,共获得6个与目标基因紧密连锁的标记(Br ID10711、Br A5~Br A9),其中Br A5与目标基因共分离,Br A9为一侧最近标记,它与目标基因之间的遗传图距为0.69 c M。至此,Brsc1基因进一步被限定于标记Y06和Br A9之间约1.2 Mb的区间内。利用本研究中获得的标记检测F2群体中3种类型单株,鉴定出一个共显性标记Br A8。将本研究中获得的SSR标记与前人研究结果进行整合,加密了Brsc1基因所在区间的标记密度。同时,特异片段与拟南芥基因组进行序列比对的结果表明,共有5个标记与拟南芥的第1染色体有较好的共线性关系,暗示Brsc1基因的同源基因可能位于拟南芥的第1染色体上。本研究中获得的标记将为Brsc1基因的克隆及利用Brsc1基因进行黄籽油菜的分子辅助育种提供有利条件。     

Selection for resistance to Plasmodiophora brassicae Wor. in oriental subspecies of Brassica rapa L.

Summary Selection for resistance to Plasmodiophora brassicae Wor. in oriental groups of Brassica rapa L.Two hundred and sixty-five cultivars of leafy, oriental bassicas were tested for resistance to 18 collections of Plasmodiophora brassicae, the causal agent of clubroot. The tests were conducted in the greenhouse at low and high level inoculum concentrations. Eleven cultivars of B. rapa pe-tsai, five cultivars of B. rapa pak-choy and three cultivars of B. rapa choy-sum consistently segregated for resistance at the lower concentration of inoculum (1000 spores/ml). All 265 cultivars were susceptible at the higher concentration (1 000 000 spores/ml). Three cultivars were used in pedigree and recurrent selection schemes for increased resistance. After three cycles of selfing resistant individuals, significantly more resistant S₃ lines were derived from each cultivar. Lines derived from two cultivars. Chinese White and PI 257236, continued to improve with each cycle of selection and demonstrated increased resistance to higher levels of inoculum (up to 1 000 000 spores/ml) New cultivars based on intercrosses of S₂ resistant individuals also had significantly better resistance than the original cultivar. After two cycles of selection in the third cultivar, PI 419007, resistance did not increase and its S₂ mass did not differ significantly from the original cultivar. Evidence that indicates resistance is pathotype-non-differential and offers an alternative to major gene, pathotype-differential types of resistance currently being introduced to the leafy oriental brassicas from other Brassica rapa groups.     

青海大黄油菜粒色性状分子标记的开发和图谱整合

利用青海大黄油菜和褐籽白菜型油菜09A-126构建BC₄和F₂分离群体, 结合AFLP与群体分离分析法(bulked segregant analysis, BSA)筛选引物, 获得5个与黄籽基因Brsc1紧密连锁的分子标记Y11~Y15。5个AFLP特异片段的序列, 均与白菜型油菜的A9染色体部分序列表现同源。将5个AFLP标记成功转化为5个SCAR标记(SC11~SC15)。利用目标基因所在染色体区段序列筛选到7个与目标基因紧密连锁的SSR标记(BrID10607、KS10760、B089L03-3和A1~A4)。利用SCAR和SSR标记扫描F₂群体中部分单株, 发现SC14和A1为共显性标记。用BC₄群体将Brsc1定位在标记Y06和A4之间1.7 Mb的区间内, 遗传距离分别为0.115 cM和0.98 cM。标记Y05和Y12与Brsc1共分离。本研究为黄籽油菜分子标记辅助选择育种体系的建立及目标基因的进一步精细定位和图位克隆奠定了基础。     

Genetic effects on biomass yield in interspecific hybrids between Brassica napus and B. rapa

This study was conducted to estimate the genetic effects on biomass yield in the interspecific hybrids between Brassica napus and B. rapa, and to evaluate the relationship between parental genetic diversity and its effect on biomass yield of interspecific hybrids. Six cultivars and lines of oilseed B. napus and 20 cultivars of oilseed B. rapa from different regions of the world were chosen to produce interspecific hybrids using NC design II. Obvious genetic differences between B. rapa and B. napus were detected by RFLP. In addition, Chinese B. rapa and European B. rapa were shown genetically differences. Plant biomass yield from these interspecific hybrids were measured at the end of flowering period. Significant differences were detected among general combining ability (GCA) effects over two years and specific combining ability (SCA) effects differences were detected in 2000. The ratios of mean squares, (σ² _GCA(f) + σ² _GCA(m)) / (σ² _GCA(f) + σ² _GCA(m) + σ² _SCA), were 89% and 88% in 1999 and 2000, respectively. This indicates that both additive effects and non-additive effects contributed to the biomass yield of interspecific hybrids and the former played more important role. Some European B. rapa had significant negative GCA effects while many of Chinese B. rapa had significant positive GCA effects, indicating that Chinese B. rapa may be a valuable source for transferring favorable genes of biomass yield to B. napus. Significant positive correlation between parental genetic distance and biomass yield of interspecific hybrids implies that larger genetic distance results in higher biomass yield for the interspecific hybrids. A way to utilize interspecific heterosis for seed yield was discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

High frequency spontaneous production of doubled haploid plants in microspore cultures of Brassica rapa ssp. chinensis

Brassica rapa (syn. Brassica campestris) ssp. chinensis is an important vegetable crop, but it is relatively recalcitrant to microspore culture. One genotype each of B. rapa ssp. chinensis var. communisand var. utilis were used formicrospore culture. Embryo production of3.8–42.4 embryos/bud was obtained. A high rate of plant regeneration directly from microspore-derived embryos without subculture was achieved by an improved protocol involving replacement of culture media and reduction of sucrose concentrations after 48 h of induction,among other modifications. More than 70%of regenerated plants were spontaneous diploids. Some spontaneous tetraploid plants were also obtained from isolated microspores of both genotypes tested. These tetraploids may be directly exploited a snew varieties in a Brassica rapabreeding programme. This revised version was published online in July 2006 with corrections to the Cover Date.     

Identification and mapping of a quantitative trait locus controlling extreme late bolting in Chinese cabbage (Brassica rapa L. ssp. pekinensis syn. campestris L.) using bulked segregant analysis

DNA markers linked to a locus controlling an extreme late bolting trait, which was originally found in a local cultivar of a non-heading leafy vegetable,‘Osaka Shirona Bansei’ (Brassica rapa L. ssp. pekinensis syn. campestris L.) were identified using bulked segregant analysis. A doubled haploid (DH) line, DH27, which is a progeny of ‘Osaka Shirona Bansei’, shows extreme late bolting, and bolts without vernalization. DH27 was crossed with a normal bolting DH line, G309. The plantlets of the parents, F₁ and F₂, were vernalized and then grown in a greenhouse. The bolting time of F₂ plants showed a continuous distribution from 19 to 231 days after vernalization (DAV), suggesting the effects of a few major genes and polygenes. Possible linkage markers for this trait were screened by modified bulked segregant analysis (BSA). The BSA using four bulks suggested that a 530-bp RAPD band RA1255C was linked to a locus controlling the bolting trait. The RAPD band was cloned and used as a probe to detect RFLP. The fragment detected a single locus, BN007-1,the segregation of which in the F₂ population matched that of RA1255C. Three other RAPDs were found to be linked to BN007-1. A quantitative trait locus(QTL) affecting the bolting time was detected around BN007-1 using MAPMAKER/QTL. Since the difference between bolting times of both the parental genotypes in the F₂ was 138 days, these markers may be useful for a marker-assisted selection (MAS) in the breeding program for late bolting or bolting-resistant cultivars in B. rapa crops. This revised version was published online in July 2006 with corrections to the Cover Date.     

Characterization of FAE1 in the zero erucic acid germplasm of Brassica rapa L.

The modification of erucic acid content in seeds is one of the major goals for quality breeding in oil-yielding Brassica species. However, few low erucic acid (LEA) resources are available, and novel LEA genetic resources are being sought. Fatty acid elongase 1 (FAE1) is the key gene that controls erucic acid synthesis. However, the mechanism for erucic acid synthesis in B. rapa lacks systematic study. Here, we isolated zero erucic acid lines from 1981 Chinese landraces of B. rapa and found that the formation of LEA is not attributable to variations in FAE1 coding sequences, as reported for B. napus, but may be attributable to the decrease in FAE1 expression. Moreover, the FAE1 promoter sequences of LEA and high erucic acid materials shared 95% similarity. Twenty-eight bases deletions (containing a 24-base AT-rich region) were identified approximately 1300 bp upstream from the FAE1 start codon in the LEA accessions. The genotype with the deletions co-segregated with the LEA trait in the segregating population. This study isolated an LEA B. rapa resource that can be exploited in Brassica cultivation. The promoter variations might modify the expression level of FAE1, and the results shed light on novel regulation mechanisms for erucic acid synthesis.     

津田芜菁BrSIZ1基因克隆、定位及表达

SIZ1是植物细胞蛋白质翻译后修饰SUMO化的E3连接酶,参与植物蛋白相互作用、定位和抗逆反应等。为研究BrSIZ1在津田芜菁中的表达特性,本研究克隆了津田芜菁SIZ1基因全长cDNA序列,命名为BrSIZ1 (GenBank登录号为KY441465),该基因全长2754 bp,其ORF全长2571 bp,编码856个氨基酸残基的多肽。构建了BrSIZ1-GFP表达载体进行亚细胞定位研究,结果显示BrSIZ1-GFP定位于细胞核内,可能在细胞核中发挥其功能。利用荧光定量PCR检测表明,该基因表达量在叶子中最高,幼苗和红色根皮中次之,表达具有组织特异性。而且BrSIZ1在芜菁根皮中的表达受长波紫外线(UV-A)诱导,在4°C、37°C胁迫的幼苗中,表达量增加。     

恰玛古脆片不同干燥方式工艺初探

旨在确定恰玛古最佳干燥工艺条件。以新疆柯坪县恰玛古为试验材料,开展了不同切分尺寸、不同前处理方式（热烫、冷冻、冷藏）试验,以及热风干燥、微波真空干燥和联合干燥试验;通过对比恰玛古在不同条件干燥过程中水分含量、水分活度、色差、膨化率、感官评定等指标的变化情况,确定最佳干燥工艺条件。结果表明,最优切分方式为片,热烫处理为6 min;冷藏处理较冷冻处理好;联合干燥方式失水速率较其他2 种方式快,水分含量、水分活度值都较低,色泽也较好,膨化率也较高。最终确定恰玛古最佳干燥工艺为：热风干燥温度45℃ 微波真空干燥1 kW 10 min,此时水分含量为7.74%、水分活度为0.19、色差L*69.4,a*4.2,b*21.0、膨化率14.80%、感官评分85。产品酥脆,色泽金黄,口感微甜。本研究具有一定的生产指导意义。     

Inter-accession variation in floral morphology in radish (Raphanus sativus L.)

We investigated variation in floral morphology among 36 accessions of radish (Raphanus sativus L.). We found wide, continuous inter-accession variation in 15 traits related to the size of the floral organs and the stigma position relative to the anthers of long stamens. A principal-component analysis of the 15 floral traits also showed wide variation among accessions. A large proportion of this variation was accounted for by lengths of stamens and the petal tube. In addition, the phenotypic stabilities with respect to the macro-environment (as affected by different sowing dates) and the micro-environment (among the flowers within a plant) differed between traits. Correlation analyses using all traits suggest the existence of genetic linkages or pleiotropies among floral organs. Our results indicate that floral morphologies varied greatly among radish accessions and they could be affected by the macro- and micro-environment. To explore the relationship between floral morphology and pollination efficiency, we compared the number of pollen grains deposited on the stigmas in the presence and absence of insect pollinators using three accessions with very different floral morphology. Because an accession with a higher stigma position than anthers might avoid self-pollination and an accession with a large stigma size received more pollen grains, floral morphology may affect pollination efficiency. Because the effect of pollination efficiency on seed production is well known, our results suggest that radish floral morphology affects seed production through its effect on pollination efficiency.     

Vegetative and floral characteristics of interspecific Brassica chimeras produced by in vitro grafting

Summary Interspecific Brassica chimeras (Brassica campestris+B. oleracea) were synthesized according to an in vitro graftculture method, and propagated by tissue culture of axillary buds or chimeric explants. A total of 127 regenerants obtained were investigated. The mericlinal chimera type was more easily produced than the other periclinal and revertant types. No sectorial chimera was produced. The flowering habit and inflorescence type of the chimeras were found to be controlled by the constitution of three tissue layers, but the petal shape and color were controlled only by the two outer tissue layers. Pollen viability of the chimeras were generally lower and more variable in parts than those of the parental types.     

Image analysis measurements and repeatability estimates of siliqua morphological traits in Brassica campestris L

Summary The repeatabilities of six traits; peduncle, pod, beak, and total siliqua length, seed number/siliqua and seed mass/siliqua were estimated for the random-mating CrGC-1 Brassica campestris L. population. All length measurements were made using a fast digital image analysis technique. Beak, pod, and total siliqua length showed the highest repeatabilities of all six traits (0.73, 0.63, and 0.63, respectively). Both pod and total siliqua length were highly correlated with both seed number and seed mass/siliqua. These results suggest that these siliqua-related morphological traits could be used as indirect indices of selection in breeding for improved seed yields in Brassica campestris L.     

密度对油菜品种机械化收获特性的影响

为了分析适宜高密度种植的油菜的重要农艺性状与产量关系的差异,为机械化高密度油菜种植提供借鉴,以不同分枝类型油菜品种为试验材料,对种植密度对油菜花期、抗倒伏性、产量的影响进行研究。结果表明,油菜的花期因为密度的增加而缩短,即开花期时间相对集中,油菜茎秆变细、茎秆抗折断力显著减弱、有效分枝的上移集中。与此同时,单位长度抗折力距呈现先增高后又降低的趋势,种植密度在44.98万株/hm2时达到最大,抗倒伏能力最强。因此,在现有栽培密度15.0万株/hm2~22.5万株/hm2的基础上,适当增加种植密度到44.98万株/hm2左右,油菜花期相对集中,缩短油菜上下部角果成熟时间上差异,茎秆单位长度抗折力距增高,更加抗倒,油菜茎秆变细、有效分枝的上移集中、株型更加紧凑,均有利于机械化收割。     

Inheritance of seed colour in Brassica campestris L. and breeding for yellow-seeded B. napus L.

Summary Seed colour inheritance was studied in five yellow-seeded and one black-seeded B. campestris accessions. Diallel crosses between the yellow-seeded types indicated that the four var. yellow sarson accessions of Indian origin had the same genotype for seed colour but were different from the Swedish yellow-seeded breeding line. Black seed colour was dominant over yellow. The segregation patterns for seed colour in F₂ (Including reciprocals) and BC₁ (backcross of F₁ to the yellow-seeded parent) indicated that the black seed colour was conditioned by a single dominant gene. Seed colour was mainly controlled by the maternal genotype but influenced by the interplay between the maternal and endosperm and/or embryonic genotypes. For developing yellow-seeded B. napus genotypes, resynthesized B. napus lines containing genes for yellow seed (Chen et al., 1988) were crossed with B. napus of yellow/brown seeds, or with yellow-seeded B. carinata. Yellow-seeded F₂ plants were found in the crosses that involved the B. napus breeding line. However, this yellow-seeded character did not breed true up to F₄. Crosses between a yellow-seeded F₃ plant and a monogenomically controlled black-seeded B. napus line of resynthesized origin revealed that the black-seeded trait in the B. alboglabra genome was possibly governed by two independently dominant genes with duplicated effect. Crossability between the resynthesized B. napus lines as female and B. carinata as male was fairly high. The sterility of the F₁ plants prevented further breeding progress for developing yellow-seeded B. napus by this strategy.     

Genetic diversity of Brassica napus L. Germplasm from China and Europe assessed by some agronomically important characters

Summary The genetic diversity and relationships among 63 rapeseed accessions, including 34 Chinese, 22 Czech, 2 Swedish, 2 German, one French and 2 Canadian accessions, were evaluated by nine agronomically important characters in the field at Yangling, Shaanxi, China. Significant differences between Chinese and European group in plant height, setting position of the first primary branch, number of siliques of the terminal raceme, thousand seed weight and seed yield per plant were detected. There were significant variations in nine agronomic characters among the tested rapeseed accessions. Ward’s minimum variance cluster analysis based on Mahalanobis distances on the raw data of nine agronomic characters clearly separated the European accessions from the Chinese ones. However, the Chinese accessions with erucic acid free and/or low glucosinolates could not be separated from those Chinese accessions with both high erucic acid and glucosinolates. In general, cluster analysis of the 63 accessions based on the selected agronomic characters was consistent with known pedigree information and geographic origin, as well as the previous RAPD results of these accessions. The European rapeseed could be important germplasm resources for enriching the genetic background of Chinese rapeseed, and vice versa.     

‘Thermally aided pollination’: A new method of breaking self-incompatibility in Brassica oleracea L.

Summary Application of heat during self-pollination of open flowers by means of an electric mini soldering iron resulted in seed set in self-incompatible Brassica oleracea varieties. Temperatures of 60, 70 and 80°C were tested. Compared with bud-pollination, in some cases the so-called thermally aided pollination (TAP) method, gave a considerably higher seed yield at 70 and 80°C. In view of the ease and rapidity of TAP it should be ascertained how far this method can replace bud-pollination in maintaining inbred lines. The possible mechanism behind the TAP response is briefly discussed.     

甘蓝型油菜小孢子培养胚发生能力的遗传分析

用 4× 4完全双列杂交研究了甘蓝型油菜小孢子培养胚状体发生能力的遗传规律。结果表明 :多数F1的胚产量与具高胚发生能力的亲本相近 ,部分F1的胚产量与双亲中亲值接近。胚状体发生能力主要由基因的加性效应控制。高胚发生能力由显性核基因控制。广义和狭义遗传力分别为 97 2 %和 81 1%。由Lisandra(高胚发生能 )×Kamikita(低胚发生能 )的F2 群体胚发生数的分离结果得出小孢子胚状体发生能力由具加性效应特点的两个基因位点控制     

Screening and identification of resistance to bacterial soft rot in Brassica rapa

Mist-chamber, field, and detached leaf inoculation procedures identified plants resistant to bacterial soft rot [Erwinia carotovora subsp. carotovora, (Ecc)] in Brassica rapa and related species. The mist-chamber seedling inoculation provided the best correlation of mean disease severity ratings with the field plant inoculation (r = 0.67^**) and was used to identify resistant materials. The optimum mist-chamber incubation conditions to distinguish the resistance of accessions were 23 ^°C and 100% RH and were used for primary screening. A total of 752 accessions of B. rapa and related Cruciferae were screened. In general, accessions of B. oleracea were more resistant than accessions of B. rapa. Within B. rapa, subspecies pekinensis and chinensis were more susceptible than other subspecies. No completely resistant material was found. In species of B. rapa, only 7% of accessions showed some degree of resistance with plant-to-plant variation within the accessions. G30444, G30449, and AVRDC2837 were identified as the most resistant materials in B. rapa by both mist-chamber and field inoculations. The resistance was correlated between an USA isolate (Geneva-1) and two Chinese isolates (RL4-1 and RL-19) of Ecc. This revised version was published online in July 2006 with corrections to the Cover Date.     

Self-incompatibility as a pollination control mechanism for spring oilseed rape,Brassica napus L.

Summary Self-incompatibility was shown to be an effective method of pollination control in spring rapeseed (B. napus L. ssp. oleifera (Metzg.)) by comparing the yield of a Westar-Topas syn-1 produced by crossing two SI lines with the yield of the corresponding syn-1 produced by hand pollination. Although the trial showed high-parent heterosis in the syn-1s, there was insufficient replication to determine the level of heterosis.Abbreviations SI self-incompatible - SC self-compatible