Effect of Combined Use of Boron and Manganese Fertilizers on the Nutritional Quality and Physiological Indices of Brassica campestris L.ssp.chinensis L.var.utilis

[Objective] The purpose of this study is to determine the effects of com-bined use of boron and manganese fertilizers on the nutritional quality and physio-logical indices of Brassica campestris. [Method] In the nutrient solutions for growing B. campestris by hydroponics, boric acid and manganese sulfate were added at 0.5, 2.5, and 7.5 mg/L respectively. Another treatment without boron and manganese was prepared as the control. Quality and physiological indices of B. campestris in the 10 treatments were measured. [Result] Boron and manganese shared obvious in-teraction in improving the quality and physiological indices of B. campestris. To cul-tivate B. campestris with high quality and strong resistance, the optimum concentra-tions of boron and manganese in the nutrient solution should be 2.5 mg/L boric acid and 2.5-7.5 mg/L manganese sulfate. [Conclusion] The findings wil provide refer-ence for studying effects of trace elements on nutrient composition of vegetables.

Preliminary Study on Tissue Culture Technique of Brassica campestris L. ssp. chinensis (L.) Makino var. utilis

In order to establish a rapid propagation system for Brassica campestris L. ssp. chinensis var. utilis, the anther and cotyledon-cotyledonary petioles were used as explants to conduct tissue culture research. The results showed that not bloomed flower buds with higher styles than petals were appropriate for anther cul- ture. Moreover, most microspores were at the mid-late uninucleate stage at that time. The pollen germination rates of Brassica campestris L. ssp. chinensis var. u- tills were low and the germination rates in autumn and winter were higher than that in summer. The induction medium of anther callus of Brassica campestris L. ssp. chinensis var. utilis was MS＋1.0 mg/L KT＋1.0 mg/L 2, 4-D＋3% sugar＋6 g/L agar＋ 8% coconut milk and the induction media of adventitious bud were MS＋2.0 mg/L 6-BA＋ 0.5 mg/L NAA＋I.0 g/L activated carbon＋2% sugar＋6 g/L agar or MS＋2.0 mg/L ZT＋ 0.5 mg/L IAA＋0.5 g/L AgNO3＋1.0 g/L activated carbon＋2% sugar＋6 g/L agar. The induction percentage of adventitious bud by anther culture was 36.7%. Browning phenomenon appeared during the culture of adventitious bud and regeneration plant could not be formed. The plant regeneration rate reached 80% when cotyledon- cotyledonary petioles were used as explant.

Role of nitrification inhibitor DMPP(3,4-dimethylpyrazole phosphate) in NO^-_3-N accumulation in greengrocery( Brassica campestris L. ssp. chinensis ) and vegetable soil

The influence of nitrification inhibitor(NI) 3,4 dimethylpyrazole phosphate(DMPP) on nitrate accumulation in greengrocery( Brassica campestris L. ssp. chinensis ) and vegetable soil at surface layer were investigated in field experiments in 2002 and 2003 Results showed that NI DMPP took no significant effect on yields of edible parts of greengrocery, but it could significantly decrease NO - 3 N concentration in greengrocery and in vegetable soil at surface layer. In addition, NI DMPP could reduce the NO - 3 N concentration during the prophase stage of storage.

不结球白菜(Brassi cacampestris ssp.chinensis Makino)种质资源SRAP遗传分化分析

利用SRAP分子标记分析了国内外64份不结球白菜种质资源的DNA遗传多样性和遗传分化。21对引物组合共检测出215个位点,其中112个为多态性位点,多态性比率达52.09%,平均每对引物组合产生10.24个位点和5.33个多态性位点。不结球白菜各类型中普通白菜的Nei’s基因多样性指数(0.1410)和遗传丰富度[(190)88.37%]最高;各生态区域中江淮流域不结球白菜的Nei’s基因多样性指数(0.1451)和遗传丰富度[(185)86.05%]最高;国内的Nei’s基因多样性指数(0.1293)和遗传丰富度[(188)87.44%]分别高于国外。遗传变异估算表明,不结球白菜遗传分化系数58.22%,大部分变异存在于种群间;基因流为0.4031,说明群体间基因流动较少,遗传分化程度较高。以遗传相似系数0.872为截值,可把不结球球白菜分为Ⅵ个类群。

Effect of Nitrogen and Sulfur Supply on Glucosinolates in Brassica campestris ssp.chinensis

Glucosinolates （GSs） are a group of plant secondary metabolites containing abundant nitrogen （N） and sulfur （S） mainly in Brassica and have the beneficial effects on human health including anti-carcinogenic, cholesterol-reducing and other pharmacological effects. The objective of this study was to investigate the effect of different concentrations of N （5, 10, and 20 mmol L-a, denoted by N5, N10 and N20） and S （0,5, 1, and 2 mmol L^-1, denoted by S0.5, S1 and S2） on the yield and GSs in pakchoi （Brassica campestris L. ssp. chinensis var. communis） in hydroponics. Results showed that N10 and N20 significantly enhanced the yield compared with N5, however, N20 had a negative effect relative to N10. Only with N10 and N20 low S supply （S0.5） reduced the yield. The concentrations of aliphatic GSs, aromatic GS and total GSs were enhanced by N5 and indolyl GSs were enhanced by N20. S2 enhanced the concentration of individual GS and total GSs. The concentrations of indolyl GSs were maximized in N20S2 treatment, whereas the highest concentrations of aliphatic GSs, aromatic GS and total GSs were found in N5S2 treatment. Effects of N and S on aliphatic GSs were higher than on indolyl GSs. The results suggest that the accumulation of aliphatic GSs and aromatic GS could be enhanced by low N and high S and restricted by high N while that of indolyl GSs could be enhanced by high N and high S.

Effect of the Antisense BcMF12 Driven by the BcA9 Promoter on Gene Silencing in Brassica campestris L.ssp.chinensis

The study analyzed the silencing of BcMF12 gene regulated by BcA9 promoter in the transgenic pakchoi and confirmed the effect of antisense BcMF12 gene on the pollen development. A conserved BcMF12 gene fragment was amplified from the cDNA of flower buds in pakchoi （Brassica campestris L. ssp. chinensis, syn. B. rapa L. ssp. chinensis） and was fused to the anther specific BcA9 promoter. The plant antisense expression vector was constructed and then introduced into pakchoi via Agrobacterium-mediated transformation. The transgenic plants were screened by antibiotics and molecular analysis. PCR and Southern blot revealed that the antisense BcMF12-GUS fusion gene regulated by BcA9 promoter was integrated into transgenic plants. Northern blot suggested that the expression of BcMF12 gene was down-regulated significantly. The pollen germination rate of transgenic plants with antisense BcMF12 gene decreased as compared with that of the control plants. The expression of the gene BcMF12 related to the pollen development was inhibited by the antisense BcMF12 driven by BcA9 promoter, which consequently affected the pollen development in pakchoi.

Effects of Different Cadmium Levels on Active Oxygen Metabolism and H_2O_2-Scavenging System in Brassica campestris L.ssp.chinensis

The effects of different Cd (Cadmium) levels on generation of active oxygen speceies(AOS) and H2O2-scavenging system in the leaves of Brassica campestris L. ssp. chinensiswere studied. The results showed that generation rate, and H2O2 content were enhancedand malondialdehyde (MDA) content increased with the increase of Cd concentrations inthe growth medium. The activities of ascorbate peroxidase (APX), dehydroascorbatereductase (DR) and glutathione reductase (GR) were promoted by the addition of Cd.Exposed to Cd also increased the contents of ascorbate (AsA) and glutathione (GSH) in theleaves.

Genetic linkage map of Brassica campestris L.using AFLP and RAPD markers

A genetic linkage map comprised of 131 loci was constructed with an F 2 population derived from an inter subspecific cross between Brassica campestris L. ssp. chinensis cv. 'aijiaohuang' and ssp. rapifera cv. 'qisihai'. The genetic map included 93 RAPD loci, 36 AFLP loci and 2 morphological loci organized into 10 main linkage groups (LGs) and 2 small groups, covering 1810.9cM with average distance between adjacent markers being approximately 13.8cM. The map is suitable for identification of molecular markers linked to important agronomic traits, QTL analysis, and even for marker assisted selection in breeding programs of Chinese cabbage and turnip.

Nitrogen Dioxide-Induced Responses in Brassica campestris Seedlings: The Role of Hydrogen Peroxide in the Modulation of Antioxidative Level and Induced Resistance

This article investigates the responses of Brassica campestris seedlings to an acute level of nitrogen dioxide （NO2） exposure in a plant growth chamber, and examines whether pretreating plants with hydrogen peroxide （H2O2） will alleviate NO2-caused injury. Twenty-eight-day-old B. campestris plants sprayed with 10 mmol L^-1 H2O2 aqueous solution （corresponding to approximate 1.0 mg H2O2 per single plant） were exposed to different concentrations of NO2 （0.25, 0.5, 1.0, and 2.0 μL L^-1, respectively） for 24 h under controlled environment. To measure the plant biomass, the plants were fumigated with the same NO2 concentrations as mentioned above for 7 h per day （8.00-15.00） for 7 days. As a control, charcoal filtered air alone was applied. Data were collected on plant biomass, total chlorophyll, photosynthetic rate, stomatal conductance, nitrate and nitrate reductase （NR）, antioxidative enzymes, ascorbate （ASA）, and malondialdehyde （MDA）, immediately after exposure. The results showed that exposure to a moderate dose of NO2 （e.g., 0.25 μL L^-1） had a favorable effect on plants, and the dry weight of the above-ground part increased, whereas the exposure to high NO2 concentrations （e.g., 0.5 μL L^-1 or higher） caused a reduction in the plant biomass and the total chlorophyll, when compared with the control. In addition, at 0.5 μL L^-1 or higher NO2 concentrations, prominent increases in the MDA level and superoxide dismutase （SOD） and NR activities were observed. Exposure to 1 μL L^-1 and higher NO2 resulted in necroses appearing on older leaves, and an increase in catalase （CAT） activity, decrease in ASA content, increased accumulation of NO3^-, and reduction in photosynthesis, when compared with the controls. No changes were detected in stomatal conductance under NO2 fumigation. The pretreatment with 10 mmol L^-1 H2O2 alleviated significantly NO2- caused biomass decrease and photosynthetic inhibition when compared with H2O2-untreated plants. Under NO2 fumigation, further induction in SOD and CAT activities occurred in H2O2 treated plants when compared with H2O2- untreated plants. The effect of NO2 on the ASA and MDA contents was also absent in H2O2-treated plants. However, the H2O2 treatment did not alter the nitrate content and NR activity in plants under NO2 fumigation. The H2O2 treatment caused a lower rate of stomatal conductance. Taken together, these data suggest that fumigation with an acute level of NO2 causes oxidative damage to B. campestris seedlings. The H2O2 pretreatment markedly protects plants against NO2 stress and this may be associated with inducible antioxidative level. NO2 fumigation contributes, at least in part, to the enhanced levels of nitrate in B. campestris leaves.

Differential expression of salt tolerance related genes in Brassica campestris L. ssp. chinensis (L.) Makino var. communis Tsen et Lee

We examined salt tolerance responsive genes in Pak-choi under salt stress and analyze their potential function. The LRNA differential display was used to screen the transcript derived fragments （TDFs） related to salinity tolerance in tolerant and Loderately tolerant Pak-choi germplasm. Seventy-eight primer combinations generated 101 differential eDNA fragments, which ere divided into 10 expression types. Seven cDNA sequences （GenBank accession Nos. DQ006915-DQ006921） obtained and ,~quenced were highly homologous to some known expression genes or the genes related to the signaling pathways in plants under ifferent abiotic stress.

白菜核雄性不育两用系AFLP扩增特异片段的克隆及序列分析

利用AFLP技术从白菜核不育两用系"矮脚黄"可育系DNA中得到一个特异扩增片段MF-14.Southern杂交结果证实不育系也存在该片段的同源序列;回收该特异扩增片段并将其克隆到pGEM-TEasy载体进行序列测定.测序结果表明,该片段全长289bp,其碱基组成为A+T=46.02%.与白菜RAPD特异片段S107及其他育性相关基因核苷酸序列比较,同源性均低于50%.表明该片段为一新的序列.

不同倍性不结球白菜Pol CMS及保持系生理生化特性比较

对二、四倍体不结球白菜Pol CMS及其保持系花蕾和薹叶进行生理生化特性比较分析,结果表明:不育系花蕾中可溶性蛋白、可溶性糖、脯氨酸含量均显著低于保持系,且各指标在不育系中四倍体低于二倍体,而在保持系中则四倍体高于二倍体;不育系MDA含量,POD、SOD、CAT活性均高于保持系,不育系中四倍体均高于二倍体,而保持系中四倍体均低于二倍体;薹叶中MDA含量,POD、SOD、CAT活性均为不育系高于保持系,不育系中四倍体高于二倍体;POD、EST同工酶显示不育系与保持系间均具特异酶带,但不同倍性间并无差异。

不结球白菜同源四倍体Pol CMS及其保持系花药发育的解剖学研究

以秋水仙素人工诱变获得的不结球白菜同源四倍体Pol CMS及其保持系为材料，采用石蜡切片法研究其花药发育过程。结果显示：四倍体Pol CMS与保持系花药发育差异明显，不育系花药发育受阻于孢原细胞分化期，不形成药室，属无花粉囊型。极少数温敏型花药能产生1～2个花粉囊，但四分体时期绒毡层提早退化，导致成熟小孢子数目减少。四倍体保持系花药发育过程与二倍体基本一致，同一时期的可育小孢子四倍体比二倍体大，二倍体花粉粒都为3个萌发孔，而四倍体多为4个萌发孔。四倍体保持系同一花粉囊中花粉粒发育有不同步现象，花粉粒败育频率比二倍体高。研究表明，不结球白菜同源四倍体Pol CMS比其二倍体Pol CMS不育更彻底，更具廊用性。

白菜转录因子BcBHLHogu的克隆及其特征分析

【目的】探明白菜'矮脚黄'OguCMS核质互作的机制。【方法】采用cDNA-AFLP技术分析白菜'矮脚黄'OguCMS及其保持系花蕾基因的表达差异,获得一个在保持系早期花蕾不表达但在OguCMS早、中、晚花蕾中稳定表达的差异片段。利用RACE技术获得其cDNA全长,命名为BcBHLHogu(GenBank登录号:EF127860)。【结果】该基因编码122个氨基酸,具有basic helix-loop-helix(bHLH)结构域;进化分析表明,BcBHLHogu与拟南芥AtBHLH060同源性最高,是冗余基因AtBHLHs060/048的直系同源基因,其功能与BEEs1/2/3(3个油菜素内酯信号的早期应答冗余基因)相似。【结论】BcBHLHogu是白菜bHLH转录因子家族的一个新成员,涉及到花器官发育信号事件,其功能与油菜素内酯应答有关。

不结球白菜BcLRK01基因对芜菁花叶病毒及胁迫诱导的响应

通过cDNA-AFLP技术,从芜菁花叶病毒(TuMV)侵染的不结球白菜幼叶中分离到一条差异表达的基因片段,克隆获得其cDNA全长为2 124bp,编码707个氨基酸的富亮氨酸重复类受体激酶,命名为BcLRK01。利用实时定量PCR研究了该基因在TuMV侵染及高盐、冷胁迫、水杨酸(SA)、茉莉酸(JA)、乙烯(ET)等处理下的表达情况,结果显示,TuMV侵染、高盐、冷胁迫、SA、JA和ET等均能诱导BcLRK01不同程度的表达,说明该基因可能是不结球白菜病毒病的病程相关基因,同时也参与高盐和冷胁迫以及SA、JA、ET等的信号途径。

菜心cDNA-AFLP分析体系的优化与建立

以菜心的顶端分生组织为试材,对影响菜心cDNA-AFLP体系的关键因子进行了探讨,以期建立适于菜心转录组差异表达的cDNA-AFLP分析体系。结果表明:Trizol法适用于菜心总RNA提取;SMART双链cDNA合成法效率较高,其中第二链cDNA合成的最佳循环数为24个;Taq I/Ase I分步各酶切3 h可使双链cDNA酶切完全;预扩增体系以模板稀释20倍,27个循环扩增效果较好,退火温度对扩增结果影响不大,但Mg2+对PCR扩增影响较大,经比较加入2.5μL Mg2+(25 mmoL/L)的预扩及选扩电泳的效果均最佳;预扩产物稀释20倍用于选择性扩增,其PAGE电泳可以获得丰富且重复性好的带型。cDNA-AFLP分析体系的优化与建立为菜心抽薹开花分子机理进一步研究奠定了基础。

CaCl_2对盐胁迫下小白菜生长和相关生理特性的影响

采用CaCl2根际注射结合叶面喷施的方法,研究了不同浓度(0、5、10、20、30 mmol/L)CaCl2对盐胁迫下小白菜(Brassica campestris ssp.chinensis L.)幼苗生长及其生理生化特性的影响。结果显示,10 mmol/LCaCl2处理显著提高了幼苗的株高、单株干重、单株鲜重和展开叶片数等,降低了叶片丙二醛(MDA)含量,增强了叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)等保护酶活性,显著提高盐胁迫下小白菜叶片的光合色素含量、净光合速率等。表明适宜浓度的外源钙可以通过增强植株的渗透调节能力和抗氧化能力,保护膜结构和功能,增强光合能力,从而减轻NaCl对小白菜的胁迫伤害。

不结球白菜细胞质雄性不育材料的筛选及其保持系选育

用多种不同基因型的可育不结球白菜(Brassica campestris ssp. chinensis Makino)材料对2个不结球白菜雄性不育株进行授粉,根据F1代群体的育性表现,筛选出细胞质雄性不育株;对16个自交品系的纯度进行调查,各品系筛选出5个或10个典型植株对其株高、开展度、叶长、叶宽、叶柄长、色泽等性状进行测试和比较分析。结果表明,16个品系的纯度均不低于50%;1-5、3-31、2-16、2-61、1-79、华冠4的开展度、株高均较大;聚类结果显示,3-31、1-9、2-61、2-16和华冠4聚在第三类,品系间性状差异较小。1-5、1-79、3-31、2-61、1-26、2-11、2-25和1-98各性状的变异系数均低于30%,一致性好;16个自交品系叶色空间分布较叶柄色集中,说明品系间叶色差异较小;3-31、1-79、1-34-17、2-11、2-16、1-5和华冠4的叶色亮丽。综合分析,1-5、1-79、3-31和2-11色泽好,变异系数小,且纯度高,是选育保持系的优良品系;而华冠4和墩子黄既可以作保持系又可以作父本。3-31和华冠4的植株高大,分别适宜作为保持系和父本,用于选配高产的不结球白菜杂交组合。

白菜GLDH基因cDNA的克隆及序列分析

以白菜品种‘苏州青’cDNA为模板,采用RT-PCR、巢式PCR、3’RACE和5’RACE技术,获得了L-半乳糖酸-1,4-内酯脱氢酶(EC1.3.2.3,GLDH)基因cDNA2034bp全长序列。序列分析表明,GLDH基因cDNA序列编码601个氨基酸。其氨基酸序列与花椰菜GLDH基因具有98%的同源性,与拟南芥GLDH基因具有90%的同源性。该基因在GenBank中登录号为AY899298。

NaCl胁迫对不同生育期红菜薹的影响

研究了NaCl胁迫对不同生育时期红菜薹[Brassica campestris L.ssp.chinensis(L.)Makino var.utilis Tsen et Lee]的影响。结果表明,在种子萌发期,300 mmol/L的NaCl处理显著降低了红菜薹种子的发芽率、发芽势和发芽指数;在幼苗期,不同浓度的NaCl处理对红菜薹的地上部鲜重、根鲜重、根干重、丙二醛含量、根系活力均无显著影响,在300 mmol/L的NaCl胁迫下红菜薹的地上部干重反而显著增加;在成株期,不同浓度的NaCl处理对红菜薹的叶片数、主薹高度、主薹茎粗、侧薹数、单株产量、丙二醛含量、根系活力均无显著影响。