大豆孢囊线虫4号生理小种侵染大豆根系诱导表达的cDNA分析

大豆孢囊线虫(Heterodera glycines Ichinohe；Soybean Cyst Nematode，SCN)是一种土传的专性内寄生线虫。SCN的二龄幼虫侵入到大豆幼嫩的根组织中，导致大豆根内的细胞变形并与之形成“合胞体”。合胞体在形态上和生理上的变化是SCN直接诱导大豆基因表达的结果。本研究以高抗SCN的灰布支黑豆为材料，用大豆孢囊线虫二龄幼虫直接接种大豆的根系，应用DDRT—PCR技术及RDB(Reversedot—blotting)杂交鉴定，获得6个阳性cDNA克隆，分别是SCN侵染后5天的A32克隆(GenBank登录号为B1173978)；侵染后10天的B12克隆(GenBank登录号为B1173979)、B71克隆(GenBank登录号为B1173980)；侵染后15天的Cll克隆(GenBank登录号为B1173981)、CPl2(GenBank登录号为B1173982)克隆和CP32克隆(GenBank登录号为B1173983)。序列的同源比较表明，6个cDNA均与Shoemaker构建的大豆基因表达库中的cDNA序列有非常高的同源性，证明这些cDNA是大豆基因表达的产物。其中A32克隆的序列与控制拟南芥下胚轴生长的MYB转录因子、营养元素缺失诱导的番茄根的表达文库中的一个cDNA及番茄抗假单胞杆菌表达文库中的一个cDNA有较高的同源性。     

大豆吡哆醇生物合成蛋白基因(PDX)的电子克隆和进化分析

电子克隆(In silicocloning)是随着基因组计划和EST计划实施而发展起来的利用生物信息学手段进行基因克隆的新方法。根据物种间同源基因相对保守的特点,以拟南芥(Arabidopsis thaliana)吡哆醇生物合成蛋白cDNA序列为信息探针,对大豆(Glycine max)EST数据库进行同源搜索和序列拼接,获得了1 280 bp长的大豆吡哆醇生物合成蛋白的基因序列(GenBank登陆号为DQ139265)。经过RT-PCR扩增、基因组PCR扩增、分子克隆和序列分析验证,结果表明与电子克隆序列完全一致。该基因具有完整的开放阅读框架(ORF,20~955 bp),编码311个氨基酸。通过与水稻、日本百脉根、烟草、截形苜蓿等物种的吡哆醇生物合成蛋白序列比对,发现该基因具有高度的保守性。表明根据物种间同源基因序列,对跨物种间EST数据库进行同源检索、筛选、拼接,是克隆基因的有效途径。     

Effect of Inoculant Rate on Nodulation and Various Agronomic Traits of Soybean

In a soil lacking indigenous Bradyrhizobium japonicum , soybean ( Glycine max [L.] Merr.) nodulation depends upon the number of rhizobia applied with the inoculum. This field study reports the effect of different rates of applied rhizobia on nodulation, dry matter and nitrogen content in soybean in a Mediterranean soil lacking B. japonicum.
Treatments included six rates of B. japonicum , ranging from 2.5 × 10⁴ to 6.075 × 10⁶ rhizobia cells per seed applied to the seed as peat inoculant at planting, 100 kg N ha⁻¹ and an uninoculated control. The experiment was conducted in an Entisol soil. Regression analysis showed linear relationship between the rate of applied rhizobia and the number of the nodules per plant or the dry weight per nodule. In early stages of development (32 and 68 days after planting) plant dry weight was not affected by inoculation rate. At harvest a rate of 7.5 × 10⁴ rhizobia cells per seed was necessary for maximum total and stover dry weight. A higher rate, 6.75 × 10⁵ rhizobia cells per seed, was required to obtain maximum grain yield, total N content in plant tops and grain N content. Grain percentage N was increased up to 2.025 × 10⁶ rhizobia cells per seed. Nitrogen application increased grain yield, total N content and grain N content at the same level as the lower inoculation rate.     

Sensitivity to Drought Stress of Three Soybean Cultivars During Different Growth Stages

Three soybean ( Glycine max L.) culivars, Lee-74, Wright , and Ra 401 , were subjected to 100 % (control), 75 %, and 50 % of field capacity during vegetative (VI), flowering (R2), and pod-filling (R4) stages in greenhouse and field studies. Stress applied at R2 significantly reduced the yield in the greenhouse, while in the field, the maximum reduction was observed when the plants were subjected to stress at R4. Stress during VI reduced the yield components less than stress applied during R2 or R4 stages in both studies. The pod number and seed weight were the yield components most affected by drought stress, and the number of seeds per pod the least affected. Yield stress index was significantly correlated with the yield and its components. The reproductive stage was clearly more sensitive to drought than the vegetative stage. The cultivar Lee 74 had the highest yield and the second highest yield stress index whereas the cultivar Ra 401 had the lowest yield.     

Identification of soybean strains resistant to Xanthomonas campestris pv. glycines

Summary Soybean germplasm was screened for resistance to bacterial pustule disease. The etiological agent, Xanthomonas campestris pv. glycines, was isolated from the leaves of field grown soybean in Maharashtra, India. The screening of soybean stocks was carried out by excised leaf inoculation method. A differential susceptibility to the pathogen was observed in the tested stocks. Two stocks P-4-2 and P-169-3 were found to be completely resistant to the pathogen and displayed an incompatible reaction. Four cultivars, EC-34160, Bragg, Kalitur and PK-472 displayed moderate resistance and the remaining stocks were susceptible to the attack of the pathogen. The stocks P-4-2 and P-169-3 remained resistant even to a high concentration of 10⁹ colony forming units (cfu)/ml of the pathogen.     

Genetic diversity in soybean as determined by RAPD and microsatellite analysis

The random amplified polymorphic DNA (RAPD) and microsatellite techniques were used to evaluate the genetic diversity among 18 soybean genotypes selected for a breeding programme to increase the protein content of varieties adapted for central European growing conditions. Out of 33 random primers used in RAPD reactions, only 12 showed polymorphism useful for characterization of these genotypes. In contrast, all 12 microsatellite primer pairs used in this study detected polymorphism with 2–6 alleles per locus. Similarity measures and cluster analysis were made using RAPD and simple sequence repeat (SSR) data, separately and together. The resulting dendrograms were compared with each other and with the available pedigree information as a control. The dendrogram derived from RAPD data showed some divergence from the pedigree information available for the lines. The dendrograms based on SSR data and SSR data combined with RAPD gave very good agreement with pedigree information. It can be concluded that the combined use of a limited number of RAPD and SSR markers is a useful and reliable means of evaluating genetic relationships of genotypes in the absence of pedigree data.     

南方菜用大豆种子越夏贮藏的研究

前言菜用大豆(Glycine max L.)俗称毛豆,我国南方普遍栽培。早熟品种如“四月拔”、“五月拔”一般在2月下旬至3月上旬播种,6月上旬开始收获。其种子在采收后要经过高温、多雨的夏季而使生活力下降。而且翌年早春播种,由于气温低和土壤湿度大,常常发生出苗慢、出苗率低,甚至烂种的现象,影响全苗和壮苗,是生产上存在的重要问题。本研究的目的,即是通过采用几种不同的简易贮藏方法比较,来寻找有效地防止菜用大豆种子越夏贮藏过程中活力下降的措施。     

Breeding strategies for early soybeans in Belarus

Yield and other agronomic traits in soybean depend on the growth habit. The best maturity group, 00 indeterminate cultivars, compared with semi determinate cultivars for 3 years, out yielded the latter by 9%, possessed greater height and node number but somewhat stronger lodging and better lower bottom pod set. Maturity did not differ significantly within the samples. Correlations were recorded between the yield, growth type and node number. Indeterminate cultivars, in general, require fewer days to start flowering than determinate cultivars. This character is useful in the development of cultivars tolerant to a moderate drought in the second half of the summer. Thus, the most likely model of maturity group 00 variety for Belarus may be an indeterminate, early flowering, full season cultivar.     

Sampling Survey and Identification of Races of Soybean Cyst Nematode (Heterodera glycines Ichinohe) in Huang-Huai Valleys

Soybean cyst nematode (SCN Heterodera glycines Ichinohe) is one of the most important nationwide soybean diseases in China. A total of 38 soil specimens or locations in the area was sampled and tested for SCN races during 2001-2003 for the inspection of race distribution in Huang-Huai Valleys. A map of race distribution was constructed according to the data from both the present study and the published reports cited. Three areas, namely, the area of southeast to Jinan in Shangdong Province; the area of northern Henan Province and its border region to south of Hebei Province; and the area of Luohe, Zhoukou of Henan Province and Fuyang of Anhui Province mainly infested with Race 1 were identified. Race 4 was predominant in Shanxi Province, Beijing and the adjacent area of Henan, Shandong, and Anhui provinces, and the delta of Huanghe River in Shandong Province. Race 2 was mainly found in Liaocheng, Dezhou of Shangdong Province and Shijiazhuang of Hebei Province, and Jiaozuo and Huojia of Henan Province. Race 7 was distributed in the west part of Jiaodong Peninsula of Shandong Province and Kaifeng, Huaxian, Wenxian of Henan Province. Race 5 was found and scattered in Hebei and Henan Province. Race 9 was found in Shangqiu of Henan Province, which was reported for the first time in China. It can be seen that Race 1 and Race 4 were the two predominant races in Huang-Huai Valleys, and that research should focus on developing resistant cultivars of these races. There might exist other races in an area with some predominant races. The race substitution in the past decade was not obviously found, therefore, the results should be meaningful to future breeding for resistance to SCN in Huang-Huai Valleys.