Research Progress of Heat-tolerance of Brassica campestris ssp. Chinensis

Brassica campestris ssp. chinensis, also known as non-heading Chinese cabbage, is an important vegetable widely distributed in southern China. High temperature is the most common adversity factor in vegetable production, because Brassica campestris ssp. chinensis is a thermophilic vegetable, which can＇t well grow at high temperature. In summer and autumn, high temperature stress would prevent Brassica campestris ssp. chinensis from growing and result in poor quality of its seedlings, seriously influencing yield and quality of the vegetable in later period. In this paper, present situation and latest advances in heat resistance research of Brassica campestris ssp. chinensis in recent years were reviewed at home and abroad. The research tendency in Bassica campestris ssp. chinensis was also described, providing reference for breeding of heat-resistant Brassica campestris ssp. chinensis.

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.

Genetic Inheritance and Molecular Marker of Clubroot Resistance Genes in Brassica campestris ssp. chinensis

Objective] This study was conducted to investigate the genetic inheritance of clubroot resistance in Chinese non-heading cabbage （Brassica campestris ssp. chinensis）. [Method] The clubroot resistance gene was introduced from a Brassica campestris ssp. pekinensis cultivar to non-heading Chinese cabbage, and the inheri-tance and molecular markers of clubroot resistance gene in parental lines, F1, F2 and BC1 of non-heading Chinese cabbage were studied through pathogen inoculation at seedling stage and ISSR-PCR. [Result] Clubroot resistance in non-heading Chi-nese cabbage was control ed by a single dominant gene. ISSR molecular markers with Bulk segregant analysis （BSA） found that primer-873 was linked to resistance gene, named CR-873, and the genetic distance between the marker and the resis-tance gene was 9.72 cM. [Conclusion] The results provide references for the molecular marker assisted breeding of non-heading Chinese cabbage.

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.

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.

Differential Expression Analysis of Genie Male Sterility A/B Lines in Chinese Cabbage-Pak-Choi (Brassica campestris ssp. chinensis Makino)

To determine differential expression of genie male sterility A/B lines in Chinese cabbage-pak-choi (Brassica campestris ssp. chinensis Makino var. communis Tsen et Lee), we used the RNA fingerprinting technique, cDNA-AFLP analysis, in different developmental stages and different tissues. While no obvious differential expressions were observed in rosette leaves, florescence leaves, and scapes, some differential expressions were found in alabstrums of A/B lines and among leaves, scapes and alabstrums. We analyzed the al-abstrums collected in different developmental stages with 10 primer combinations. We got a unique band between middle size alabstrums and large alabstrums in B line in one of the ten pair primers, and in another one pair, one band reflecting a higher gene-expression level in A line than that in B line was obtained. No unique bands were found with the other primer combinations. The bands reflecting different gene-expression level were confirmed by Northern hybridization. The results indicated that cDNA-AFLP was a suitable tool for studying differential expression of genie male sterility in plants. SDS-polyacrylamide gel electrophoresis patterns of soluble proteins further verified the difference in A/B lines.

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.

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.

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.

A Genetic Linkage Map of Brassica campestris L.ssp. pekinensis (syn. B. rapa L. ssp. pekinensis)

A molecular genetic map of Chinese cabbage was constructed with a 102 recombinant inbred (RI) population from a cross of two cultivated Chinese cabbage lines 177 and 276, using AFLP and RAPD markers. 352 markers including 265 AFLP markers and 87 RAPD markers were integrated into 17 linkage groups. It covered a total of 2 665. 7 cM with an average interval of 7. 6 cM. AFLP marker is efficient for map construction while it easily forms clusters to cause big gaps in map. A total of 13.92 % abnormal segregation markers distributed in the map. The molecular genetic map is fundamental for gene localization, comparative genomics, and QTL mapping of important agronomic traits.

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。

铬胁迫对青菜(Brassica chinensis L.)植物螯合肽含量及抗氧化特性的影响

选用不同耐铬胁迫青菜品种春油1号和青伏令,采用不同浓度Cr3+处理,测定青菜抗氧化酶活性及植物螯合肽含量等生理指标。结果表明,随着Cr3+处理浓度升高,两种青菜叶片抗氧化酶活性、酸溶性SH、植物螯合肽(PCs)及谷胱甘肽(GSH)含量均呈增加趋势(P<0.05),高浓度Cr3(+300 mg.L-1)处理下,耐铬胁迫较弱的青伏令叶片SOD、POD、CAT活性分别比对照增加了24.2%、41.7%、21.4%,耐铬胁迫较强的春油1号分别比对照增加了37.2%、70.7%、43.0%,并且PCs含量是对照的3.17倍,明显高于耐铬胁迫较弱品种青伏令(1.71倍)。Cr3+处理下耐铬胁迫较强的青菜品种春油1号表现出相对较高的抗氧化酶活力及非酶物质含量,不同品种青菜对铬胁迫的生理差异较大。

铜、锌污染对水稻土中油菜(Brassica chinensis L.)生长的影响及累积效应研究

通过盆栽试验,系统研究了Cu、Zn重金属污染对油菜(Brassica chinensis L.)生长、SOD保护酶活性、脯氨酸、可溶性糖及Cu、Zn在油菜体内吸收富集的影响。结果表明:Cu、Zn污染对油菜生物量和生理生化指标(SOD保护酶活性、脯氨酸和可溶性糖)均有影响。随着Cu、Zn质量分数的升高,植株干物重和SOD保护酶活性表现为先增后降;当Cu质量分数达到1 000 mg?kg-1时,油菜干物重和SOD酶活性分别比对照下降36.79%、66.67%;当Zn质量分数达到3 000 mg?kg-1时,油菜干物重和SOD酶活性分别比对照下降55.21%、82.05%。Cu污染下,油菜叶片脯氨酸质量分数逐渐下降,总可溶性糖质量分数表现为先增后降;Zn污染下,脯氨酸和总可溶性糖质量分数均表现为先增后降。Cu、Zn污染使油菜幼苗叶片及根中的Cu、Zn质量分数明显增加,根部的积累尤其明显。当Cu处理为1 000 mg?kg-1时,油菜茎叶和根中质量分数分别为57.6、424.162 mg?kg-1;当Zn处理为1 000 mg?kg-1时,油菜茎叶和根中质量分数分别为240、608.54 mg?kg-1。与Zn相比,Cu主要积累在油菜的根部,向茎叶迁移累积的量很少,并且随着添加质量分数的增加,茎叶吸收Cu的量变化不大。总之,重金属Zn相对于Cu更容易积累于油菜的地上部分,从而更易进入食物链,它们在油菜体内富集都对其生理生化和营养产生了明显毒害作用。

镉胁迫对小白菜(Brassica campestris L.)抗氧化机理的影响

采用水培的方法,研究了不同Cd2+水平(0、1、2.5、5、10、15mg·L-1)对小白菜(Brassica campestris L.)抗氧化酶(SOD、POD、CAT)活性、非酶物质(SH、GSH、PCs)含量以及生长的影响。结果表明,小白菜叶片和根系SOD活性随Cd处理质量浓度的增加呈降低的变化趋势,POD、CAT活性以及MDA含量随Cd处理质量浓度的增加而增加。小白菜的生物量、根长、株高、叶绿素质量分数随Cd处理质量浓度的增加显著降低(P<0.05),表明Cd抑制了小白菜的生长,破坏了叶绿素的合成。小白菜地上、地下部镉质量分数均随Cd处理质量浓度的增加而显著增加(P<0.05)。当Cd处理质量浓度为15mg·L-1,小白菜地上、地下部镉质量分数分别达到637.5、1663.0mg·kg-1,表明小白菜对Cd有良好的富集效果。小白菜根系与叶片中SH、GSH和PCs含量均随Cd处理质量浓度增加而增加的变化趋势,表明SH、GSH和PCs在解毒小白菜Cd毒害中起着重要作用。

沼液对无土栽培小白菜(Brassica chinensis L.)产量及品质的影响初探

无土栽培条件下,初步研究了沼液和化学肥料组合施用对小白菜(Brassica chinensis L.)产量及品质的影响。结果表明,当所施用的沼液营养液肥配方为每升沼液营养液肥中含沼液250 mL,配入3.0g尿素、2.7 g磷酸二氢钾、1.1 g氯化钙、2.5 g硫酸镁和2 mL微量元素储备液(处理3),小白菜产量最高,为10.6克/株,而且,与单纯施用化学肥料营养液(对照)相比,小白菜品质指标硝氮、Vc、可溶性糖及叶绿素含量并没有下降。生产中,合理利用沼液栽培小白菜,对提高其产量和品质有积极的作用。

鸡毛菜(Brassica rapa L.Chinensis Group.)的颜色变化动力学及两种货架期预测方法

实验测定了贮藏在278、283、288、293K四个温度下的鸡毛菜的颜色参数L*、-a*、b*、△E、h*和感官评价,并对L*和△E进行了动力学分析。结果表明,在实验温度范围内,鸡毛菜储藏的温度越低,颜色参数变化越慢。动力学分析显示,零级动力学比一级动力学更适合表现鸡毛菜L*和△E的变化规律。研究还采用Arrhenius方程对颜色变化速率常数k和温度T进行非线性拟合,得到L*和△E活化能Ea分别为82.09kJ/mol和100.26kJ/mol。最后得到依赖于时间、温度和颜色指标的鸡毛菜货架期预测方程(R2>0.95),从中根据不同的L*和△E终点可得到对应的货架期。就本研究而言,假定L*增加12%为终点时,鸡毛菜在四个温度下的预测货架期分别为11.8、5.5、2.6、1.3d。同时,Arrhenius方程与依赖于感官终点的动态颜色终点拟合方程结合预测的货架期曲线与感官寿命曲线则能得到更好的契合(两者绝对差值小于0.6d),两者结合能得到较为全面的货架期预测参数。

Study on Agrobacterium tumefaciens-mediated Transformation of Brassica campestris L. with Fusion Gene Ycoil-bFGF

[ Objective] The study is to generate pharmaceutical protein via plant transgenic technique. [Methed] Using the cotyledons with petiole as transformation receptor, the fusion gene of rapeseed oil-body gene and bFGF was introduced into the rapeseed （ Brassica campestris L. ） by Agrobacterium tumefaciens-mediated transformation; meanwhile regeneration conditions of rapeseed were also optimized, and the regenerated resistant plantlets were detected by PCR and Southern blot. [ Result] This fusion gene had been integrated into rapeseed genome successfully, and the optimized conditions of transformation and regeneration were as follows： explants pre-culture for 2 d, co-culture for 3 d, bacteria solution OD600 for 0.3 and infection time for 5 min. [ Conclusion] The results laid a solid foundation for extraction, isolation and purification of protein in transgenic plant seeds.

Effects of field application of phosphate fertilizers on the availability and uptake of lead, zinc and cadmium by cabbage (Brassica chinensis L.) in a mining tailing contaminated soil

A field trial was conducted to evaluate the reduction of bioavailability of heavy metals including lead (Pb), zinc (Zn) and cadmium (Cd) in a soil contaminated by mining tailings in Shaoxing, Zhejiang, China. Three commercial phosphate (P) fertilizers including phosphate rock (PR), calcium magnesium phosphate (CMP), and single superphosphate (SSP) were applied to the plot at three P application rates, 50, 300, and 500 g/m2 with 9 treatments and control (CK). Plants, water soluble and exchangeable (WE) extra...

Analysis on Cloning and Expression of WRKY Gene Segment in Brassica campestris L.

[Objective]The aim was to explore the relative expression trends of WRKY transcription factor gene under ABA treatment in Brassica campestris.[Method]Actin gene and WRKY gene were cloned by using the homology cloning method.The sequences of nucleic acid and amino acid were analyzed using BLAST and DNAMAN software.Relative expression trends of WRKY gene were detected by applying real-time relative quantification PCR（RT-qPCR）under ABA（100 μmol/L）treatment.[Result]A 680 bp WRKY gene segment and a 933 bp β-actin gene segment were acquired in Brassica campestris.The result of RT-qPCR analysis revealed that BcWRKY expression could be induced by ABA and that the relative expression of WRKY gene reached the peak at 1 h with ABA treatment.[Conclusion]Actin and WRKY gene in B.campestris were cloned successfully,which was proved to play an important role in ABA signal pathway.

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.