Multivariate Based Variability within Diverse Indian Mustard (<i>Brassica juncea</i>L.) Genotypes

Indian mustard (Brassica juncea L.) germplasm consisting 167 accessions including one check cultivar was evaluated for qualitative and quantitative traits. The present study was conducted to investigate genetic diversity and correlation among studied genotypes of B. juncea L. based on agro-morphological at NARC, Islamabad, Pakistan. To investigate the genetic diversity based on morphological characters, data was recorded on 20 quantitative and 12 qualitative traits. The calculated data was analyzed through two complementary methods, i.e. PCA (Principal Component Analysis) and cluster analysis. Among all the studied cultivars, significant diversity was recorded for different agro-morphological characters. Among all the parameters, maximum variance was recorded for pod shattering (427.2) followed by plant height (345.6), days to 100% flowering (336.2) and main raceme length (210.0). Among all the characters, the greatest and highly significant association (0.99) was found between days to maturity 50% and days to maturity 100% followed by correlation (0.86) among days to flowering 50% and days to flowering 100%, correlation value (0.71) was calculated among leaf length and leaf width. Using cluster analysis all the genotypes were divided into five major groups. It was observed that 7 out of 20 principal components with an Eigen value of ≥1.0 calculated for 73.92% of the total diversity observed between 167 accessions of Indian mustard (B. juncea L.). The contribution of first three PCs in the total PCs was 23.25, 12.87 and 11.24, respectively. Among all the investigated accessions two genotypes 26,813 and 26,817 showed great potential for seeds/silique, 1000-seed weight and seed yield/plant, respectively, so these genotypes are recommended for future breeding programs for achieving promising results.

芥菜(Brassica juncea L.)小孢子胚发生和植株再生

对芥菜(Brassica juncea L.)的7种基因型进行游离小孢子培养试验,从供体母株取2～4mm长的花蕾,分离小孢子并将其置于NLN-82液体培养基中薄层培养,先在33℃,后移到25℃继续暗培养,3周后统计形成的球形期至子叶期胚.将成熟的小孢子胚转移到无激素MS琼脂培养基直接或经二次分化得到小孢子植株.7种基因型均观察到细胞分裂,6种基因型获得了小孢子胚,3种基因型得到了再生植株.各基因型小孢子胚胎发生频率差异很大;产量最高的“成都大头菜”,平均每蕾小孢子胚产量为10.0个.

三种类型油菜(Brassica spp.)和野芥菜(B.juncea var.gracilis Tsen et Lee)杂交亲和性及F_1的适合度——潜在基因转移的研究

通过人工去雄授粉 ,采用荧光显微镜观察了 3种类型栽培油菜花粉在两地采集的野芥菜柱头上的萌发生长情况 ,结合杂交后的结实率 ,探讨了 3种类型油菜和野芥菜杂交的亲和性 .结果表明 ,甘蓝型油菜和芥菜型油菜与野芥菜的亲和性都非常高 ,亲和性指数达 10 .0以上 ,而白菜型油菜和野芥菜的亲和性较低 ,亲和指数小于 0 .2 .子一代的适合度研究结果表明 ,芥菜型油菜向野芥菜杂交一代的适合度没有降低 ,而甘蓝型及白菜型和野芥菜杂交一代的适合度明显下降 ,表现在花粉活力降低 ,结实率极低 .上述结果表明 ,白菜型油菜和野芥菜的基因转移可能性最小 ,甘蓝型居中 ,而芥菜型极易向野芥菜发生基因转移 .表 4参

Differentially Expressed Genes Related to Fertility Conversion in Thermo-sensitive Genic Male Sterile(TGMS) Lines of Brassica juncea

[Objective] This study was performed to screen functional genes related to the fertility conversion of thermo-sensitive genic male sterile （TGMS） lines of Brassica juncea L. [Method] A B. juncea TGMS line K121S was selected as the experimental material. The total RNAs were isolated from fertile and sterile pollens at different development stages, including mother cell stage, tetrad stage, tricellular pollen stage and maturity stage. DDRT-PCR was carried out to identify differentially expressed genes. [Result] A total of 44 differentially expressed cDNA fragments were identified with Dot blot. And seven candidate genes related to fertility conversion of K121S were screened out by BLASTN, including callose synthase gene, aldehyde dehydrogenase gene and RNA polymerase I transcription factor RRN3 gene which were differentially expressed at the transcriptional level, H＇-ATPase gene, fructose diphosphate aldolase -class I gene, teucine-rich repeat receptor-Jike serine/threonine- protein kinase gene and alkaline/neutral invertase gene, which were differentially expressed at the post-transcriptional level. [Conclusion] The results of this study will help to explain the molecular mechanism of thermo-sensitive genic male sterility of B. juncea.

Variation and Statistic Analysis of Agronomic Traits of Recombinant Inbred Lines of Brassica juncea L.

Correlation analysis and principal component analysis( PCA) were conducted for some agronomic traits of 139 recombinant inbred lines of Brassica juncea L. The results showed that under the environmental conditions in Guiyang,the flowering time,number of seeds per silique,plant height and thousand seed weight differed significantly among different lines. The results of principal component analysis showed that the lines could be classified into three groups. The results of Pearson correlation tests showed that the plant height was positively correlated with the flowering time( P < 0. 05),and the thousand seed weight was negatively correlated with the flowering time and number of seeds per silique( P < 0. 01).

体积排阻高效液相色谱-电感耦合等离子体质谱法同时测定印度芥菜(Brassica Juncea)中镉铜锌形态

建立了体积排阻高效液相色谱-电感耦合等离子体质谱法同时测定印度芥菜中镉、铜、锌形态分析方法。为了防止巯基化合物氧化反应的发生,样品采取液氮保护并在-70℃保存,样品分析全流程采取氮吹防氧化措施。在镉、铜、锌胁迫下,诱导产生植物螯合肽(PCs)。在叶片和根部均检测到3种元素的4种形态,植物螯合肽(PC)3-Cd(Cu,Zn)、植物螯合肽(PC)2-Cd(Cu,Zn)、谷胱甘肽(GSH)-Cd(Cu,Zn)、半胱胺酸(Cys)-Cd(Cu,Zn)及其在植物体内的分布规律。结合植物不同部位Cd、Cu、Zn分布规律初步推断,Cd、Cu、Zn在与GSH及Cys的结合上存在竞争。

Nitrogen in Relation to Photosynthetic Capacity and Accumulation of Osmoprotectant and Nutrients in Brassica Genotypes Grown Under Salt Stress

Different strategies of the application of nutrients are required to overcome the adverse effects of mustard （Brassica juncea L.） in response to NaCl stress. The objective of the present study was to determine if different added levels of nitrogen （N） in growth medium could alleviate the adverse effects of salt stress on photosynthetic capacity and accumulation of osmoprotectants and nutrients. 14 days mustard seedlings of salt-sensitive （cv. Chuutki） and salt-tolerant （cv. Radha） genotypes were fed with： （i） 0 mmol L^-1 NaCl ＋ 0 mg N kg^-1 sand （control）, （ii） 90 mmol L^-1 NaCl ＋ 30 mg N kg^-1 sand, （iii） 90 mmol L^-1 NaCl ＋60 mg N kg^-1 sand, （iv） 90 mmol L^-1 NaCl ＋90 mg N kg^-1 sand and （v） 90 mmol L^-1NaCl＋ 120 mg N kg^-1 sand. Under the condition of salinity stress, N application caused a significant ameliorative effect on both genotypes with respect to growth attributes [fresh weight （FW） and dry weight （DW）] and physio-biochemical parameters [percent water content （WC）, net photosynthetic rate （PN）, stomatal conductance （gs）, total chlorophyll （Ch1）, carbonic anhydrase （CA） activity and malondialdehyde （MDA）, nitrogen （N）, potassium （K） and sodium （Na） contents, and K/Na ratio] and yield attributes （number of pods/plant, seeds/pod and seed yield/plant）. The salt-tolerant genotype exhibited maximum value for growth, physio-biochemical and yield attributes at 60 mg N kg 1 sand than that of salt-sensitive genotype. These results suggest that application of N may ameliorate most of the attributes and prove to be a physiological remedy to increase the tolerance against the ill effects of salt stress in Brassicas.

芥菜型油菜每角籽粒数QTL的上位性互作和环境互作分析

【目的】为揭示芥菜型油菜及芸薹属作物每角籽粒数形成的分子机理,提高和改良芥菜型油菜产量和育种工作奠定基础。【方法】研究以包含221个芥菜型油菜株系的重组自交系(recombinant inbred line,RIL)群体为材料,在5个环境条件下对每角籽粒数性状进行加性QTL、加性×加性上位互作及环境互作分析。【结果】(1)共检测到7个与每角籽粒数相关的加性QTL,主要分布在芥菜型油菜A02、A03、A05、A08、B02和B03等染色体上,其加性效应分布在(-11.6424)~4.5246之间,其中qSS2-71的加性效应和遗传率均最大,分别达到-11.6424和14.44%,其余6个加性QTL的加性效应和遗传率均较小;(2)检测到7对影响每角籽粒数的加性×加性QTL上位互作效应及其与环境的互作效应,上位性QTL互作效应值分布在(-4.9308)~4.1936之间,7对上位性QTL与不同环境互作的遗传力均接近0;(3)每角籽粒数性状的广义遗传率为80.98%,狭义遗传率为30.98%。【结论】综合分析,芥菜型油菜每角籽粒数受一定环境影响,但控制该性状的加性效应受环境影响较小,且其加性×加性上位性QTL互作效应不明显。

用茎瘤芥再生芽离体筛选抗S－（2－氨乙基）－L－半胱氨酸变异体研究

将茎瘤芥“棒棒菜”（Ｂｒａｓｓｉｃａｊｕｎｃｅａｖａｒ．ｔｕｍｉｄａｃｖ″Ｂａｎｇｂａｎｇｃａｉ″）下胚轴离休培养的再生芽，用１２０～４０Ｇｙγ射线照射后，在原培养基（ＭＳ＋ＮＡＡ０．２ｍｇ／ｌ＋ＢＡ２．０ｍｇ／ｌ）上培养１周，转到加Ｓ－（２－氨乙基）－Ｌ－半胱氨酸（ＡＥＣ）的原培养基中，经过连续７代选择，获得了抗ＡＥＣ的变异体植株。其蛋氨酸和精氨酸含量分别为对照的１．８４倍和３倍，赖氨酸比对照略有增加，总氨基酸含量为对照的１．４５倍。

芥菜型油菜黄化突变体L638-y的遗传及黄化基因gr1的分子标记

【目的】研究芥菜型油菜黄化突变体L638-y黄化性状的遗传规律,并初步定位黄化基因gr1。【方法】以芥菜型油菜黄化突变体L638-y分别与正常绿色的渭源大黄芥、2598进行杂交,根据杂交F1、F2和BC1材料单株的叶色调查结果进行遗传分析;以从L638-y与2598杂交后代选育得到的近等基因系(Near-isogenic line,NIL)为材料,选用SRAP、SSR、RAPD和AFLP分子标记,采用BSA(Bulk segregant analysis)法,初步定位黄化基因gr1。【结果】芥菜型油菜黄化突变体L638-y的黄化性状受2对隐性核基因控制,将其分别命名为gr1、gr2。将黄化基因gr1初步定位在2个AFLP分子标记EA4TG4和EA7MC1之间,其与2个标记间的遗传距离分别为33.6与21.5cM。【结论】芥菜型油菜黄化突变体L638-y的黄化性状受2对隐性核基因控制,且黄化基因gr1初步定位在EA4TG4和EA7MC1之间。

Assessment of Genetic Variation Within Indian Mustard (Brassica juncea) Germplasm Using Random Amplified Polymorphic DNA Markers

Genetic diversity among 45 Indian mustard （Brassica juncea L.） genotypes comprising 37 germplasm collections, five advance breeding lines and three improved cultivars was investigated at the DNA level using the random amplified polymorphic DNA （RAPD） technique. Fifteen primers used generated a total of 92 RAPD fragments, of which 81 （88%） were polymorphic. Of these, 13 were unique to accession ‘Pak85559＇. Each primer produced four to nine amplified products with an average of 6.13 bands per primer. Based on pairwise comparisons of RAPD amplification products, Nei and Li＇s similarity coefficients were calculated to evaluate the relationships among the accessions. Pairwise similarity indices were higher among the oilseed accessions and cultivars showing narrow ranges of 0.77-0.99. An unweighted pair-group method with arithmetic averages cluster analysis based on these genetic similarities placed most of the collections and oilseed cultivars close to each other, showing a low level of polymorphism between the accessions used. However, the clusters formed by oilseed collections and cultivars were comparatively distinct from that of advanced breeding lines. Genetically, all of the accessions were classified into a few major groups and a number of individual accessions. Advanced breeding lines were relatively divergent from the rest of the accessions and formed independent clusters. Clustering of the accessions did not show any pattern of association between the RAPD markers and the collection sites. A low level of genetic variability of oilseed mustard was attributed to the selection for similar traits and horticultural uses. Perhaps close parentage of these accessions further contributed towards their little diversity. The study demonstrated that RAPD is a simple and fast technique to compare the genetic relationship and pattern of variation among the gene pool of this crop.

白芥子与黄芥子UPLC特征图谱差异性研究

目的:研究白芥子与黄芥子饮片、水煎液、配方颗粒特征图谱的差异性。方法:UPLC法测定白芥子与黄芥子饮片、水煎液、配方颗粒特征图谱,采用"中药色谱指纹图谱相似度评价系统(2012版)"对色谱图进行匹配,并对特征图谱结果进行化学模式识别研究。结果:分别建立了白芥子与黄芥子饮片、水煎液、配方颗粒的UPLC特征图谱。白芥子饮片特征图谱确定了13个共有峰,黄芥子饮片特征图谱确定了11个共有峰,二者均有11个共有峰可在对应的水煎液和配方颗粒中找到;聚类分析及主成分分析结果显示白芥子与黄芥子的饮片、水煎液、配方颗粒UPLC特征图谱差异显著。结论:建立的UPLC特征图谱能够全面有效地区分白芥子与黄芥子的饮片、水煎液、配方颗粒,为芥子配方颗粒的基原鉴定、提取生产过程质量控制以及成品质量标准制定提供依据。

芥菜SRO基因家族全基因组鉴定与表达分析

【目的】SRO基因家族是植物特有的一类转录因子,在植物生长发育与胁迫响应中具有重要作用,研究芥菜SRO基因家族,为解析芥菜SRO基因功能与遗传改良提供理论依据。【方法】利用生物信息学鉴定油用芥菜与菜用芥菜基因组中的SRO基因家族成员,运用TBtools、MEGA、Cytoscape、NCBI、STRING、EggNOG等软件与数据库进行理化性质、序列特征、进化关系、调控网络等分析以及RT-qPCR分析盐胁迫下的表达模式。【结果】两种类型芥菜SRO基因家族成员数量存在差异,并分属A与B两大类,其中A类SRO蛋白含有WWE、PARP及RST结构域,而B类蛋白则缺少WWE结构域,SRO基因启动子区域含有多种非生物胁迫响应与激素响应的顺式作用元件,microRNA-BjuSRO-靶基因构建复杂调控网络,参与了细胞凋亡、根系形态建成、免疫应答、异源刺激的细胞反应、对活性氧的反应等生物学过程,油用芥菜BjuOSRO基因与甘蓝SRO基因具有较近的进化关系,RT-qPCR结果显示,在盐胁迫下BjuVA1a、BjuVA1e、BjuVA2a、BjuVA3a、BjuVB1b、BjuVA2a显著上调表达。【结论】芥菜SRO基因家族具有功能多样性,BjuVA1a、BjuVA1e、BjuVA2a、BjuVA3a、BjuVB1b、BjuVA2a与盐胁迫响应密切相关,可作为培育耐盐型芥菜新品种的候选基因。

潮州咸菜发酵工艺优化及其品质研究

文章以包心芥菜和南姜为原料,采用自然发酵制备潮州咸菜,并探究了其pH、亚硝酸盐含量、氨基酸含量和硬度等品质,重点研究了食盐用量、南姜用量和腌制时间对产品感官评分的影响。结果表明,在最佳条件下(食盐用量为65 g,南姜用量为130 g,腌制时间为7 d),产品的感官评分最高,达到85.63分,且产品中亚硝酸盐含量为0.22 mg/kg,远低于食品安全国家标准中规定的限量标准20 mg/kg。另外,产品的色泽呈现黄绿色且明暗适中,整体质地变软,爽脆程度整体下降,并且产品中的甜味与鲜味氨基酸含量较高,能够产生良好的风味。该研究对潮州咸菜的制备具有一定的参考价值,有利于潮州特色农产品包心芥菜的加工发展。

Bioactivity of methanolic extract of Brassica juncea in animal model of diabetes mellitus

Objective:To study the bioactivity of methanolic extract of Brassica juncea on animal model of diabetes mellitus along with its effect on diabetic and metabolic parameters.Methods:Diabetes mellitus was induced in rats by injecting streptozotocin(60 mg/kg)intraperitonealy.Blood glucose was measured on day 3 by GOD-POD method to confirm the diabetes mellitus.Rats having fasting blood glucose>250 mg/d L were further selected for study and they were divided into four groups,control,control+streptozotocin,streptozotocin+metformin(75 mg/kg)and streptozotocin+extract of B.juncea(450 mg/kg).Each group consisted of six rats of either sex.Metformin and experimental extract were administered for 21 d.Triglyceride,cholesterol level were measured on day 21 by commercially available kit.Blood glucose was measured on days 7 and 21.Anti-oxidant potential was assessed by estimating extent of lipid peroxidation(LPO)by malondialdehyde(MDA),nitric oxide(NO),superoxide dismutase(SOD)and glutathione(GSH)in liver,kidney,pancreas,muscle tissues on day 21.Unpaired and paired student’s t-test was applied for statistical analysis.Results:The extract of B.juncea showed significant decrease in blood glucose level on day 21.The treatment group showed significant difference in oxidative stress by increasing SOD and GSH and decreasing LPO and NO activity on day 21.The treatment did not show statistically significant difference of cholesterol,and triglycerides level on day 21.Conclusion:The study showed anti-hyperglycemic and anti-oxidative properties of methanolic extract of B.juncea.

不同施肥处理对镉污染土壤雪里蕻光合特性的影响

通过盆栽试验研究了Cd污染土壤不同施肥处理对雪里蕻光合特性的影响,以期通过有效的农业措施降低土壤重金属污染对人类健康的影响。结果表明:不同施肥处理显著提高了雪里蕻的叶绿素含量和对光照强度的适应范围,使净光合速率(Pn)、光饱和点(LSP)和表观量子效率(AQY)显著升高,使光补偿点(LCP)降低。施用有机肥可以保持较高的Pn、LSP和AQY,而且与有机肥的施用方式有关。增施磷肥使雪里蕻的Pn和LSP最大。各施肥处理下Fv/Fm的值均大于0.8,表明雪里蕻未受到光抑制。可见,适宜的施肥措施可以改善Cd污染土壤雪里蕻的光合特性,从而促进其生长。

淡紫拟青霉A10的促生特性及其发酵液对印度芥菜幼苗蓄积Cs^+的影响

从种植印度芥菜的盆栽土壤(ρ[Cs^+]为10 mmol·kg-1)中筛选出淡紫拟青霉(A10),通过测试分析A10溶磷、解钾、分泌植物生长素(IAA)类似物及其发酵液对印度芥菜幼苗蓄积Cs^+的影响,研究A10的植物促生特性及其发酵液在印度芥菜蓄积Cs^+过程中的作用,为植物-微生物互作在Cs^+污染环境修复中的应用及其互作机制研究提供实验依据。结果显示:A10对Cs^+的耐受性和富集能力强,ρ(Cs^+)为50～100 mg·L^(-1)时A10的溶磷能力被显著抑制(P<0.05);Cs^+处理及外源色氨酸均可显著诱导A10分泌IAA类似物;用1/2 Hoagland营养液稀释A10发酵液后再配制ρ(Cs^+)为25～100 mg·L^(-1)的处理液,用其培养印度芥菜,ρ(Cs^+)为0 mg·L^(-1)(CK)时A10发酵液对印度芥菜幼苗根的生长呈现"低促高抑"的生长素效应,稀释35倍时的促根效果最佳;A10发酵液稀释5、35倍和55倍条件下,ρ(Cs^+)为100 mg·L^(-1)时幼苗地上部Cs^+蓄积量均显著升高,其中稀释35倍时Cs^+蓄积量达最大值,地上部为8.86 mg·g-1DW,地下部为16.76 mg·g-1DW,印度芥菜幼苗蓄积Cs^+与其吸收K+呈显著的负相关性。研究表明,淡紫拟青霉A10具有显著的植物促生特性,A10发酵液有利于提高印度芥菜幼苗对外源Cs^+的蓄积。

陕北黄芥黄色种皮的遗传研究

采用黄芥(Brassica juncea L.)与两个不同来源的褐籽类型芥菜型油菜杂交,研究黄芥黄色种皮的遗传。结果表明:黄芥的粒色主要受母体基因型控制,褐色对黄色为显性;F2和BC1世代株间乃至株内颜色不尽一致,但黄色单株和褐色单株能够明显区分,褐色和黄色的比例分别符合3∶1和1∶1的分离比例,证明黄芥的粒色主要受一对主效基因控制,同时受修饰基因和环境的影响。黄籽和褐籽杂交F2粒色与几个品质性状的相关分析,结果证明除含油量与粒色密切相关外,其脂肪酸组分的平均值与种皮颜色也有一定的相关性。

EDDS对印度芥菜Cu积累的影响及与P-ATPase的关系研究

通过水培的方法研究了可降解螯合剂EDDS对印度芥菜对Cu积累的影响,分析了根系中与Cu主动排出有关的P型ATPase的活性及其基因表达。结果表明:根系中Cu的积累量随外界浓度的升高而升高,但Cu在茎杆与叶片中的积累量则变化不大。随着外加EDDS浓度的增加,根、茎、叶中的Cu的积累量均出现下降的趋势。上述结果表明,根系以Cu离子为主要的吸收形式,而不是EDDS-Cu。通过对印度芥菜根系细胞膜上P型ATPase活性及基因表达分析后发现,不加EDDS时,随着外界Cu浓度的升高,ATPase活性增强,而当Cu达到16μmol L-1时出现降低的趋势。但是,ATPase活性随着外加EDDS的浓度增加而降低,这是因为EDDS降低了外界的Cu活度。RT-PCR分析结果表明,该ATPase的基因(BjHMA)在转录水平上的表达与活性变化一致。由于P型ATPase的作用是将Cu排出细胞,因此上述结果说明,其活性大小以及转录水平的变化受到外界Cu离子活度的影响,在一定的范围内可以调节植物对Cu的积累。

印度芥菜对Cu诱导的氧化胁迫响应

研究了印度芥菜对不同浓度 (0、 2、 4、 8和 1 6 μmol·L-1 )Cu处理的响应。结果表明 ,4～ 1 6 μmol·L-1 Cu抑制印度芥菜根和地上部生物量的积累 ,植株中H2 O2 和丙二醛 (MDA)含量上升 ;根和叶中超氧物歧化酶 (SOD ,EC1 1 5 1 1 )和过氧化物酶 (POD ,EC1 1 1 1 7)活性均随着Cu浓度增加而上升 ;2～ 8μmol·L-1 Cu能提高根中抗坏血酸过氧化物酶 (APX ,EC1 1 1 1 1 1 )活性 ,叶中APX活性最高时的外界Cu浓度为 4 μmol·L-1 ;过氧化氢酶 (CAT ,EC1 1 1 1 6 )活性在 2 μmol·L-1 Cu时最高 ;Cu浓度为 2～ 8μmol·L-1 时根中非蛋白巯基化合物 (NPT)