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     

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     

Mechanisms of methyl-carbon transfer from S-methylcysteine to pectin in Chinese cabbage (Brassica pekinensis Rupr.)

In order to learn how the methyl group of S-methylcysteine wasmetabolized and its carbon was incorporated into the methylester of pectin in Chinese cabbage, leaves were fed S-methylcysteinewhich was labeled in the methyl group with both ³H and ¹⁴C.Incorporation of the radioactive isotopes into S-adenosylmethioninewas detected with little reduction in the ³H/¹⁴C ratio betweenthe methyl groups. The changes in the ³H/¹⁴C ratio between thepectin methyl ester recovered from the leaves and the S-methylcysteinefed to them indicate that there are at least two pathways inthe transfer of the methyl-carbon from S-methylcysteine to themethyl ester of pectin: one is the intact methyl group transfer,probably through S-adenosylmethionine, and the other is carbontransfer after the degradation of the methyl group. Cysteinesulfoxide lyase (EC 4.4.1.4 [EC] ) was found in the leaves and rootsin Chinese cabbage and its involvement in the methyl-carbontransfer is discussed. (Received December 8, 1975; )     

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.     

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.     

三分体形成是大白菜2n雄配子发生的主要途径

用能自然产生２ｎ配子的二倍体大白菜ＢＰ０５８为材料,研究了２ｎ雄配子发生的细胞学机制。结果表明,大白菜２ｎ雄配子的形成主要是由于减数分裂过程中,中期Ⅱ两个纺锤体的定向发生改变所致,即由正常的相互垂直定向改变为八字形和平行形定向。     

硅对受土壤中镉污染的白菜生长和抗胁迫能力的影响

在土培盆栽条件下,硅可提高白菜的地上部鲜重和茎鲜重,但根鲜重下降,叶鲜重略有下降。硅可抑制白菜吸收镉,在0.3、0.6、1.2mg·kg~(-1)镉水平下,施硅可显著降低白菜地上部的镉含量,并在一定程度上提高叶中过氧化物酶(POD)活性,但降低超氧化物歧化酶(SOD)活性。0.5、1.0 g·kg~(-1)硅可提高白菜的叶绿素含量和过氧化氢酶(CAT)活性,降低叶细胞膜透性,从而提高其对镉胁迫的耐受力。较高的硅浓度对植物有一定的毒性,叶绿素含量和CAT活性都下降,细胞膜透性也增加。     

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.     

Conditions controlling relative uptake of potassium and rubidium by plants from soils

Earlier studies have demonstrated close inverse relationships between Rb⁺ concentrations in plants and pH or base (including K⁺) saturation of soils. This study aims at elucidating conditions in soils influencing plant uptake of Rb⁺. Growth experiments with Carex pilulifera L. were performed, modifying the acidity and K⁺ supply of acid soils and solutions. We were unable to assess any reduction in Rb⁺ uptake by adding precipitated CaCO₃ to acid soil unless pH was raised to near neutrality. Though not fully compensating the loss of soil solution K⁺and exchangeable K⁺ from uptake by the growing plants, soil treated with 0.5 mM K⁺ (as KCl) reduced the Rb⁺ concentration in the shoots by 40% without measurably changing soil pH. Experiments varying the pH and K⁺ concentration of a nutrient solution (20% Hoagland), spiked with 6 uM Rb⁺, clearly demonstrated that plant uptake of Rb⁺ and K⁺ was unaffected by acidity in the pH range 3.6–5.0 tested, whereas Rb⁺ uptake was reduced by ca. 50%, when K⁺ concentration was increased from 1.2 to 3.6 mM. The sensitivity of this reaction indicates that shortage or low availability of K⁺ controls Rb⁺ uptake from acid soils, being probably more important than soil acidity per se. Secondary effects of high soil acidity, such as leaching losses of K⁺, might also be of importance in accounting for the high uptake of Rb⁺ from such soils. It is suggested that leaf analysis of Rb⁺ may be used as a method to assess early stages of K⁺ deficiency in plants on acid soils.     

Utilization of the bar gene to develop an efficient method for detection of the pollen-mediated gene flow in Chinese cabbage (Brassica rapa spp. pekinensis)

To develop an efficient screening method for detection of the transgene in Chinese cabbage (Brassica rapa spp. pekinensis) utilizing Basta spray, optimal conditions for Basta application were examined in this study. Two transgenic Chinese cabbage lines were obtained through Agrobacterium-mediated transformation and used as transgenic positive controls in the Basta screening experiment. Differential concentrations of glufosinate-ammonium were sprayed into three different growth stages of 12 commercial Chinese cabbage cultivars. The results showed that no plants could survive higher than 0.05% glufosinate-ammonium, and plants at the 2–3 leaf stage were most vulnerable to glufosinate-ammonium. On the other hand, no damage was observed in the transgenic control plants. Reliability of the Basta spray method was proven by showing perfect co-segregation of the tolerance to glufosinate-ammonium and the presence of the bar gene in T₁ segregating populations of the transgenic lines, as revealed by both PCR and Southern blot analyses. Using the developed Basta screening method, we tried to investigate the transgene flow through pollen dispersal, but failed to detect any transgene-containing non-transgenic Chinese cabbages whose parents had been planted adjacent to transgenic Chinese cabbages in field conditions. However, the transgene was successfully detected using Basta spray from the non-transgenic plants bearing the transgene introduced by hand-pollination. Since the Basta spray method developed in this study is easy to apply and economical, it will be a valuable tool for understanding the mechanism of gene flow through pollen transfer and for establishing a biosafety test protocol for genetically modified (GM) Chinese cabbage cultivars.     

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。     

PlantGM: a database for genetic markers in rice (Oryza sativa) and Chinese cabbage (Brassica rapa)

The Plant Genetic Map Database (PlantGM) has been developed as a web-based system which provides information about genetic markers in rice (Oryza sativa) and Chinese cabbage (Brassica rapa). The database has three major parts and functions; (1) Map Search, (2) Marker Search, and (3) QTL Search. At present, the database provides characterization information for about 3258 genetic markers. It has 2800 RFLP and 112 QTL markers related to rice in addition to 321 RFLP and 25 PCR-based markers for Chinese cabbage. In addition, a genetic linkage map was also constructed by using 1,054 markers from 2,912 markers in rice.

Availability

The database is available for free at http://www.niab.go.kr/nabic/PlantGM     

Physiological effects of Na2SO4 and NaCl on callus cultures of Brassica campestris (Chinese cabbage)

Hypocotyl-derived callus cultures of Brassica campestris L. ssp. pekinensis cv. Kim-jung (Chinese cabbage) were grown on Murashige and Skoog medium containing no additional salt, NaCl or Na₂SO₄. Na₂SO₄ was more than twice as inhibitory in comparison to the same concentration of NaCl when growth and fresh:dry weight ratios of established callus were measured. Levels of protein, starch, sucrose and α-amino nitrogen were not significantly altered in salt-grown callus. Concentrations of reducing sugars and chlorophyll were 2–3 times greater in callus grown on either salt. Proline concentration increased 15–20 fold on the highest levels of salt. Final concentrations (reached in 20–24 days) were closely correlated to the initial Na⁺ concentration of the medium, regardless of salt type. The osmotic potential in callus transferred to NaCl or Na₂SO₄ reached a maximum negative value after 16 days. For both salts, subsequent increases were correlated to increases in fresh:dry weight and growth. On both salts, turgor remained relatively constant (0. 6–0.75 MPa). Changes in Na⁺, K⁺, Mg²⁺ and Ca²⁺ content were correlated to initial Na⁺ concentration in the medium, not salt type. Accumulation of Na⁺ was accompanied by loss of K⁺ and Mg²⁺. Six to seven times less sulfate was measured in callus grown on Na₂SO₄ than chloride in callus grown on similar concentrations of NaCl.     

The construction of a genetic linkage map of non-heading Chinese cabbage （Brassica campestris ssp. chinensis Makino）

Non-heading Chinese cabbage （Brassica carnpestris ssp. chinensis Makino） is one of the most important vegetables in eastern China. A genetic linkage map was constructed using 127 doubled haploid （DH） lines, and the DH population was derived from a commercial hybrid ＂Hanxiao＂ （lines SW-13 x L-118）. Out of the 614 polyrnorphic markers, 43.49% were not assigned to any of the linkage groups （LGs）. Chi-square tests showed that 42.67% markers were distorted from expected Mendelian segregation ratios, and the direction of distorted segregation was mainly toward the paternal parent L-118. After sequentially removing the markers that had an interval distance smaller than 1 cM from the upper marker, the overall quality of the linkage map was increased. Two hundred and sixty-eight molecular markers were mapped into 10 LGs, which were anchored to the corresponding chromosome of the B. rapa reference map based on com- mon simple sequence repeat （SSR） markers. The map covers 973.38 cM of the genome and the average interval distance between markers was 3.63 cM. The number of markers on each LG ranged from 18 （R08） to 64 （R07）, with an average interval distance within a single LG from 1.70 cM （R07） to 6.71 cM （R06）. Among these mapped markers, 169 were sequence-related amplified polymorphism （SRAP） molecular markers, 50 were SSR markers and 49 were random amplification polymorphic DNA （RAPD） markers. With further saturation to the LG9 the current map offers a genetic tool for loci analysis for important agronomic traits.     

大白菜细胞质雄性不育系RAPD特异扩增片段的克隆及其序列分析

利用RAPD技术从大白菜细胞质雄性不育系CMS3411-7的DNA中得到1个特异扩增片段CMSO-PL01670,进一步以该片段为探针进行Dot和Southern杂交证实了该片段为不育系所特有。回收该特异扩增片段并将其克隆到pGEM-T Easy载体上进行序列测定。测序结果表明该片段全长673bp,其碱基组成为A T=67.16％。通过与GenBank EMBL DDBJ PDB中的455 972个序列进行比较,同源性均小于30％,表明该片段为一新发现序列。并且该序列的 4 - 216的区域内含有1个可编码70个氨基酸的开放阅读框架。上述结果为今后从分子水平进一步研究大白菜细胞质雄性不育打下了坚实的基础。