BrpSPL9 (Brassica rapa ssp. pekinensis SPL9) controls the earliness of heading time in Chinese cabbage

The leafy heads of cabbage (Brassica oleracea), Chinese cabbage (Brassica rapa ssp. pekinensis), Brussels sprouts (B. oleracea ssp. gemmifera) and lettuce (Lactuca sativa) comprise extremely incurved leaves that are edible vegetable products. The heading time is important for high quality and yield of these crops. Here, we report that BrpSPL9‐2 (B. rapa ssp. pekinensis SQUAMOSA PROMOTER BINDING‐LIKE 9‐2), a target gene of microRNA brp‐miR156, controls the heading time of Chinese cabbage. Quantitative measurements of leaf shapes, sizes, colour and curvature indicated that heading is a late adult phase of vegetative growth. During the vegetative period, miR156 levels gradually decreased from the seedling stage to the heading one, whereas BrpSPL9‐2 and BrpSPL15‐1 mRNAs increased progressively and reached the highest levels at the heading stage. Overexpression of a mutated miR156‐resistant form of BrpSPL9‐2 caused the significant earliness of heading, concurrent with shortening of the seedling and rosette stages. By contrast, overexpression of miR156 delayed the folding time, concomitant with prolongation of the seedling and rosette stages. Morphological analysis reveals that the significant earliness of heading in the transgenic plants overexpressing BrpSPL9‐2 gene was produced because the juvenile phase was absent and the early adult phase shortened, whereas the significant delay of folding in the transgenic plants overexpressing Brp‐MIR156a was due to prolongation of the juvenile and early adult phases. Thus, miR156 and BrpSPL9 genes are potentially important for genetic improvement of earliness of Chinese cabbage and other crops.     

Agrobacterium-mediated transformation of cotyledonary explants of Chinese cabbage (Brassica campestris L. ssp. pekinensis)

 A procedure for producing transgenic Chinese cabbage plants by inoculating cotyledonary explants with Agrobacterium tumefaciens strain EHA101 carrying a binary vector pIG121Hm, which contains kanamycin-resistance and hygromycin-resistance genes and the GUS reporter gene, is described. Infection was most effective (highest infection frequency) when explants were infected with Agrobacterium for 15 min and co-cultivated for 3 days in co-cultivation medium at pH 5.2 supplemented with 10 mg/l acetosyringone. Transgenic plants of all three cultivars used were obtained with frequencies of 1.6–2.7% when the explants were regenerated in shoot regeneration medium solidified with 1.6% agar. A histochemical GUS assay and PCR and Southern blot analyses confirmed that transformation had occurred. Genetic analysis of T1 progeny showed that the transgenes were inherited in a Mendelian fashion. Received: 15 December 1998 / Revision received: 2 July 1999 · Accepted: 8 July 1999     

春化温度对大白菜花芽分化和抽薹的影响

以冬性有明显差异的 6个大白菜品种为试材 ,研究了 4个春化温度对大白菜的花芽分化和抽薹的影响 .结果表明 ,弱冬性材料在 3～ 11℃范围内 ,花芽分化和抽薹均能在较短的时间完成 ;强冬性材料适宜的春化温度为 3～ 7℃ ,超过 7℃ ,花芽分化和抽薹急剧延迟 ;冬性中等材料适宜的春化温度为 3～ 9℃ .故一般采用 3～ 7℃的春化温度对不同冬性材料均适宜     

Medium and genotype factors influencing shoot regeneration from cotyledonary explants of Chinese cabbage (Brassica campestris L. ssp. pekinensis)

Medium conditions for reliable shoot regeneration from cotyledonary explants of Chinese cabbage were examined. Maximum shoot regeneration was obtained in the presence of 5 mg/l BA and 0.5 mg/l NAA. Shoot induction was further improved by the addition of AgNO₃ as well as higher concentrations (1.2–1.6%) of agar in the regeneration medium. When 123 genotypes were tested, a large variation in regeneration frequency was observed, ranging from 95% to 0%. Shoot regeneration frequency was not related to origin and days to maturity of the genotypes. Ethylene production from cultured explants seemed to play an important role in shoot regeneration. Explants of highly responsive genotypes or if cultured on the medium solidified with a higher concentration of agar generally showed low levels of ethylene production. However, AgNO₃, which also enhanced shoot induction, resulted in an increase in ethylene production. The possible interaction between ethylene and shoot regeneration is discussed. Received: 26 September 1997 / Revision received: 6 March 1998 / Accepted: 20 March 1998     

Effects of molybdenum on ascorbate-glutathione cycle metabolism in Chinese cabbage (Brassica campestris L. ssp. pekinensis)

A hydroponic trial was conducted to investigate effects of molybdenum (Mo) on ascorbate-glutathione cycle (AsA-GSH cycle) metabolism in Chinese cabbage (Brassica campestris L. ssp. pekinensis). Mo was applied at four rates: 0, 0.01, 0.15 and 1.5 mg l⁻¹. The concentrations of ascorbate, dehydroascorbate, reduced- and oxidized- glutathione, and activities of five key enzymes in the AsA-GSH cycle were studied. The results showed that appropriate Mo application increased the fresh weight of Chinese cabbage, but excess application of Mo (1.5 mg l⁻¹ Mo) decreased the fresh weight. Total ascorbate and reduced ascorbate concentrations in the Chinese cabbage increased with Mo application rates. Although no significant differences existed in DHA concentration between the different Mo regimes, but it has an increase trend with the 0.01 mg l^-1 Mo treatment, and then decreased with the Mo level increasing. No significant difference in GSH concentration was found between the different Mo treatments. Compared with the control, the GSSG concentration decreased significantly in the 0.01 mg l⁻¹ Mo treatment. The activities of APX, MDHAR, DHAR and GR increased due to Mo application. But the activity of AAO decreased with increasing Mo application rates. It is hypothesized that Mo may promote the redox process and regeneration of ascorbic acid, and affect the ability of anti-oxidation in the Chinese cabbage. Responsible Editor: Jian Feng Ma.     

Characterization of a cDNA encoding cysteine proteinase inhibitor from Chinese cabbage (Brassica campestris L. ssp. pekinensis) flower buds

A cDNA encoding a new phytocystatin isotype named BCPI-1 was isolated from a cDNA library of Chinese cabbage flower buds. The BCPI-1 clone encodes 199 amino acids resulting in a protein much larger than other known phytocystatins. BCPI-1 has an unusually long C-terminus. A BCPI-1 fusion protein expressed in Escherichia coli strongly inhibits the enzymatic activity of papain, a cysteine proteinase. Genomic Southern blot analysis revealed that the BCPI gene is a member of a small multi-gene family in Chinese cabbage. Northern blot analysis showed that it is differentially expressed in the flower bud, leaf and root.     

Erwinia carotovora ssp.carotovora Infection Induced“Defense Lignin”Accumulation and Lignin Biosynthetic Gene Expression in Chinese Cabbage(Brassica rapa L.ssp.pekinensis)

Erwinia carotovora subsp. carotovora (Ecc) infects and causes soft rot disease in hundreds of crop species includingvegetables, flowers and fruits. Lignin biosynthesis has been implicated in defensive reactions to injury and pathogeninfection in plants. In this work, variations of lignin content and gene expression in the molecular interaction betweenChinese cabbage and Ecc were investigated. H_2O_2 accumulation and peroxidase activity were detected by 3, 3-Dimethoxybenzidine staining at mocked and Ecc-inoculated sites of Chinese cabbage leafstalks. Mason lignin contentin inoculated plants increased by about 7.84%, 40.37%, and 43.13% more than that of the mocked site at 12, 24 and 72 hafter inoculation, respectively. Gas chromatography detected more p-coumaryl (H) and less coniferyl (G) and sinapyl (S)monolignins in leafstalks of Chinese cabbage. All three monomers increased in Ecc-infected leafstalks, and the Ecc-induced"defense lignin" were composed of more G and H monolignins, and less S monolignin. After searching the expressedsequence tags (EST) data of Chinese cabbage, 12 genes putatively encoding enzymes involved in lignin biosynthesis wereselected to study their expression. All of these genes could be induced by mock inoculation and Ecc infection, while thegene expression lasted for several more hours in the infected samples than in mocked and untreated plants. Our resultsindicated that "defense lignin" was different from the developmental lignin in composition; G and S monolignins weresignificantly induced in plants in response to the soft rot Ecc; thus, lignin biosynthesis was differentially regulated andplayed a role in plant response to the soft rot Ecc.     

Erwinia carotovora ssp. carotovora Infection Induced ＂Defense Lignin＂ Accumulation and Lignin Biosynthetic Gene Expression in Chinese Cabbage （Brassica rapa L. ssp. pekinensis）

Erwinia carotovora subsp, carotovora （Ecc） infects and causes soft rot disease in hundreds of crop species including vegetables, flowers and fruits. Lignin biosynthesis has been implicated in defensive reactions to injury and pathogen infection in plants. In this work, variations of lignin content and gene expression in the molecular interaction between Chinese cabbage and Ecc were investigated. H2O2 accumulation and peroxidase activity were detected by 3, 3＇- Dimethoxybenzidine staining at mocked and Ecc-inoculated sites of Chinese cabbage leafstalks. Klason lignin content in inoculated plants increased by about 7.84%, 40.37%, and 43.13% more than that of the mocked site at 12, 24 and 72 h after inoculation, respectively. Gas chromatography detected more p-coumaryl （H） and less coniferyl （G） and sinapyl （S） monolignins in leafstalks of Chinese cabbage. All three monomers increased in Ecc-infected leafstalks, and the Ecc-induced ＂defense lignin＂ were composed of more G and H monolignins, and less S monolignin. After searching the expressed sequence tags （EST） data of Chinese cabbage, 12 genes putatively encoding enzymes involved in lignin biosynthesis were selected to study their expression. All of these genes could be induced by mock inoculation and Ecc infection, while the gene expression lasted for several more hours in the infected samples than in mocked and untreated plants. Our results indicated that ＂defense lignin＂ was different from the developmental lignin in composition; G and S monolignins were significantly induced in plants in response to the soft rot Ecc; thus, lignin biosynthesis was differentially regulated and played a role in plant response to the soft rot Ecc.     

大白菜雄性不育系的组织培养和快速繁殖技术研究

以大白菜雄性不育系Bm5-3为材料,研究其组培快繁技术的结果表明:腋芽以组培快繁技术保存、扩繁大白菜雄性不育系是可行的。适合其腋芽增殖的培养基是MS+2mg·L-16-BA+0.1mg·L-1NAA,增殖系数为4.58,一般通过4￣5次继代即可脱分化;适合其生根的培养基是B5+0.1mg·L-1IBA,平均每株生根数为8.5条,平均根长8.18cm,这一培养基上生长的根较粗壮。具有4~5片叶子且根系发达的试管苗,移栽成活率高达95%。田间栽培的试管苗与供体植株的外部形态与生理特性完全一致。     

Molecular characterization of novel haplotypes of eIF4E family in Chinese cabbage (Brassica rapa L. ssp. pekinensis)

Two genes coding for eukaryotic translation initiation factors, eIF4E.a and eIF4E.c, were isolated from twelve accessions of Chinese cabbage (Brassica rapa L. ssp. pekinensis). Polymorphism analysis revealed that 94 and 142 polymorphic sites were characterized from allele of BraeIF4E.a and BraeIF4E.c which produced complex haplotype structures. Six novel haplotypes were characterized from the two alleles respectively. Among the six novel haplotypes of BraeIF4E.a, three loss-of-function mutations were identified in which a conserved single nucleotide deletion mutation cause the early termination of BraeIF4E.a coding product; while for six new BraeIF4E.c haplotypes, their coding product show amino acid substitution mutations on non-conservative amino acid residues which might affect TuMV infection in Chinese cabbage.     

不同形态微生物菌剂对不结球白菜生长和品质的影响

合理施用微生物菌剂可显著改善蔬菜土壤环境,增加产量,提升品质.本试验以不结球白菜为供试材料,在设施条件下设置不施微生物菌、施用液态和固态微生物菌剂处理,研究不同形态微生物菌剂对不结球白菜生长和品质的影响.结果表明:施用液态和固态微生物菌剂均能显著提高土壤脲酶活性,增加植株全氮含量,增大叶面积,显著提升叶片SPAD值,提...     

与大白菜霜霉病抗性主效QTL连锁的分子标记开发

霜霉病是危害大白菜的三大病害之一,该病的发生会严重影响大白菜的产量及品质,因而研究与霜霉病抗性QTL紧密连锁的分子标记对大白菜抗病新品种培育具有重要意义。该研究在前期工作的基础上,选用高感霜霉病株系91-112、高抗霜霉病株系T12-19以及由二者为双亲构建的DH群体为实验材料,针对大白菜霜霉病抗性主效QTL——BrDW所在的标记区间,利用已有的大白菜基因组信息发展与抗性QTL紧密连锁的分子标记,通过Blast和IMap分析,将与BrDW连锁的RAPD标记K14-1030定位于大白菜KBrB058M10上(位于Contig214上),根据KBrB058M10附近的BAC及BAC-end序列设计引物,结合限制性内切酶酶切及HRM分析方法,筛选得到5个与BrDW连锁的分子标记,包括1个Indel标记Brb062-Indel230,3个CAPS标记Brb094-DraⅠ787、Brb094-AatⅡ666和Brb043-BglⅡ715,1个SNP标记Brh019-SNP137;同时,通过筛选与目标区域具有同源性的Unigene序列得到了1个与BrDW紧密连锁的SSR标记bru1209。标记Brb062-Indel230、Brb094-DraⅠ787、Brb094-AatⅡ666、Brb043-BglⅡ715、Brh019-SNP137和bru1209与RAPD标记K14-1030之间的遗传距离分别为4.3 cM、1.7 cM、5.9 cM、5.9 cM、4.6 cM和0.8 cM,在对DH群体中的抗性株系选择上准确率分别为69.7%、70.9%、72.4%、72.4%、58.3%和74.2%,可应用于分子标记辅助选择,为霜霉病抗性分子育种奠定了良好基础。