外源质粒（基因）导入花生根瘤菌的行为分析

利用二亲本或三亲本杂交的方法,将携带有共生固氮基因的外源重组质粒或外源载体质粒导入慢生型花生根瘤菌［Ｂｒａｄｙｒｈｚｏｂｉｕｍｓｐ．（Ａｒａｃｈｉｓ）］１４７－３和快生型花生根瘤菌［Ｒｈｉｚｏｂｉｕｍｓｐ．（Ａｒａｃｈｉｓ）］８５－７中。探讨了转移接合子中外源质粒在人工培养条件下和共生条件下的稳定性,发现外源质粒在花生根瘤菌中的稳定性与质粒的类型、受体菌的特性和环境条件有关。同时还探讨了外源质粒上的共生基因对受体菌１４７－３共生固氮效率的影响。结果表明,外源共生基因对共生固氮能力的影响是复杂的,既可以产生正效应,也可以产生负效应。     

大豆的农艺性状与大豆灰斑病抗性关系的初步分析

对大豆灰斑病抗性水平不同的品种资源的农艺性状,包括结荚习性、株型、叶形、花色、茸毛色进行调查分析,结果表明,在灰斑病抗性较好的品种中,分枝型、圆叶、紫花、棕色茸毛的品种所占比率较大,这种现象可供今后灰斑病抗源筛选及抗灰斑病育种参考。     

大豆的农艺性状与大豆灰斑病抗性关系的初步分析

对大豆灰斑病抗性水平不同的品种资源的农艺性状,包括结荚习性、株型、叶形、花色、茸毛色进行调查分析,结果表明,在灰斑病抗性较好的品种中,分枝型、圆叶、紫花、棕色茸毛的品种所占比率较大,这种现象可供今后灰斑病抗源筛选及抗灰斑病育种参考.     

导入dctABD和parCBA／DE基因提高大豆慢生根瘤菌固氮效率和稳定性的研究

以pLAFR3为载体构建重组质粒pHN207,携带有来自苜蓿根瘤菌（Sinorhizobium meliloti）的四碳二羧酸转移酶基因dctABD、来自pTR102的parCBA／DE基因和标记发光酶基因luxAB。利用2亲本杂交法,将重组质粒pHN207导入大豆慢生根瘤菌（Bradyrhizobium japonicum）TA11和CB1809,分别考察了转移接合子中外源重组质粒在人工培养条件和共生条件下的稳定性,结果表明par基因的引入明显提高pLAFR3在TA11和CB1809中的稳定性。dctABD基因可显著提高TA11和CB1809在大豆黑龙33、宁镇一号和渝豆一号上的共生固氮能力,使结瘤植物的地上部分干重（生物量）和总氮量等指标较对照组有显著提高。     

大豆抗灰斑病菌多个生理小种资源的筛选

用人工接种大豆灰斑病茼的方法对224份供试材料,在生理小种水平上进行了抗性鉴定,筛选出抗8个以上生理小种的多抗资源47份,占供试材料的20．98％,其中东北农业大学的材料最多,比例达46．81％;黑龙江省主栽品种有5份,占多抗材料的10．64％。供试材料中,有抗单个小种和抗多个甚至全部生理小种的品种,表明在现行大豆品种资源中,具有较丰富的专化抗性资源和广谱性的抗源。     

大豆抗灰斑病主基因的发现与遗传研究

利用高抗品种东农9674与感病品种杂交,在田间多个生理小种共存条件下研究大豆灰斑病抗性的遗传规律,发现杂交后代的抗性表现具有明显的质量性状遗传特征,F1代表现完全显性,F2代的抗感分离比例在个别组合接近3:1。采用数量性状的主要基因-多基因混合遗传模型对抗性的遗传进行模型的判别与遗传参数的估计,发现抗性遗传存在明显的主要因效应,分别符合一个主基因 多基因加显性模型及两个基因独立遗传模型。主基因的加性、显性以及主基因之间的相互作用普遍存在,对抗病性的遗传起很大作用。     

Evaluation of indigenous bacterial strains for biocontrol of the frogeye leaf spot of soya bean caused by Cercospora sojina

Aims: Assessment of biological control of Cercospora sojina, causal agent of frogeye leaf spot (FLS) of soya bean, using three indigenous bacterial strains, BNM297 (Pseudomonas fluorescens), BNM340 and BNM122 (Bacillus amyloliquefaciens). Methods and Results: From cultures of each bacterial strain, cell suspensions and cell‐free supernatants were obtained and assayed to determine their antifungal activity against C. sojina. Both mycelial growth and spore germination in vitro were more strongly inhibited by bacterial cell suspensions than by cell‐free supernatants. The Bacillus strains BNM122 and BNM340 inhibited the fungal growth to a similar degree (I～52–53%), while cells from P. fluorescens BNM297 caused a lesser reduction (I～32–34%) in the fungus colony diameter. The foliar application of the two Bacillus strains on soya bean seedlings, under greenhouse conditions, significantly reduced the disease severity with respect to control soya bean seedlings and those sprayed with BNM297. This last bacterial strain was not effective in controlling FLS in vivo. Conclusions: Our data demonstrate that the application of antagonistic bacteria may be a promising and environmentally friendly alternative to control the FLS of soya bean. Significance and Impact of the Study: To our knowledge, this is the first report of biological control of C. sojina by using native Bacillus strains.     

Fine Mapping and Characterization of Candidate Genes that Control Resistance to Cercospora sojina K. Hara in Two Soybean Germplasm Accessions

Frogeye leaf spot (FLS), caused by the fungus Cercospora sojina K. Hara, may cause a significant yield loss to soybean growers in regions with a warm and humid climate. Two soybean accessions, PI 594891 and PI 594774, were identified to carry a high level of resistance similar to that conditioned by the Rcs3 gene in ''Davis''. Previously, we reported that the resistance to FLS in these two plant introductions (PIs) was controlled by a novel gene (s) on chromosome 13 that is different from Rcs3. To fine-map the novel FLS resistance gene(s) in these two PIs, F_{2: 3} seeds from the crosses between PI 594891 and PI 594774, and the FLS susceptible genotype ''Blackhawk'' were genotyped with SNP markers that were designed based on the SoySNP50k iSelect BeadChip data to identify recombinant events and locate candidate genes. Analysis of lines possessing key recombination events helped narrow down the FLS-resistance genomic region in PI 594891 from 3.3 Mb to a 72.6 kb region with five annotated genes. The resistance gene in PI 594774 was fine-mapped into a 540 kb region that encompasses the 72.6 kb region found in PI 594891. Sequencing five candidate genes in PI 594891 identified three genes that have several mutations in the promoter, intron, 5'', and 3'' UTR regions. qPCR analysis showed a difference in expression levels of these genes in both lines compared to Blackhawk in the presence of C. sojina. Based on phenotype, genotype and haplotype analysis results, these two soybean accessions might carry different resistance alleles of the same gene or two different gene(s). The identified SNPs were used to develop Kompetitive Allele Specific PCR (KASP) assays to detect the resistance alleles on chromosome 13 from the two PIs for marker-assisted selection.     

大豆疫霉根腐病抗源筛选

由大豆疫霉菌引起的大豆疫霉根腐病是大豆生产的重要病害,该病已在我国大豆主要产区发生,并在局部地区造成较大产量损失。利用抗病品种是防治大豆疫霉根腐病最有效的方法。本研究目的是筛选大豆疫霉根腐病抗源,为病害防治和抗病品种的选育提供参考。用下胚轴创伤接种方法对120个栽培大豆品种（系）进行接种,鉴定其对10个具有不同毒力大豆疫霉菌菌株的抗性。有110个品种（系）分别抗1～10个大豆疫霉菌菌株,其中以河南大豆品种（系）对疫霉菌的抗性最丰富,安徽、湖北和山西大豆品种（系）也具有抗性多样性。120个大豆品种（系）对10个大豆疫霉菌菌株共产生57个反应型,有4个抗性反应型分别与单个抗病基因的反应型一致,有7个抗性反应型与2个已知基因组合的反应型相同,其他抗性反应型为新的类型。一些大豆品种（系）中可能存在有效的抗大豆疫霉根腐病新基因。     

大豆种质资源对灰斑病抗性鉴定评价

近年来,随着大豆品种和气候的变化,灰斑病在黑龙江大豆产区广泛发生,并呈逐年加重的趋势。为了筛选出可在生产或育种中利用的抗病材料,本试验在人工接种条件下,对1073份大豆材料进行抗灰斑病鉴定和评价,结果表明,高抗材料33份、抗病材料291份、中抗材料332份、感病材料381份、高感材料36份,分别占供试材料的3.1%、27.1%、30.9%、35.5%、3.4%。对1、6、7号出现频率较高的优势生理小种鉴定结果是高抗和抗病材料分别是161份、68份、95份,分别占供试材料的15%、6.3%、8.9%。在现行大豆种质资源中,具有比较丰富的抗病资源。     

A Model Integrating Components of Rate-reducing Resistance to Cercospora Leaf Spot in Sugar Beet

Four components of rate-reducing resistance to Cercospora leaf spot in sugar beet (infection efficiency of conidia RC₁, incubation period RC₂, size of necrotic spots RC₃ and spore yield RC₄), previously measured in single infection cycle experiments, were integrated into a model simulating the chain of infection cycles under field conditions, as influenced by weather. To integrate resistance components, variables accounting for infection frequency, incubation period, affected leaf area, and infectiousness – which are computed for a susceptible cultivar – were modified by means of coefficients which reduced (RC₁, RC₃, RC₄) or increased (RC₂) them. Outputs obtained by running the model and changing resistance components actually reduced the rate of disease progress and the area under the disease progress curve of epidemics (AUDPC), as happens at field level; therefore, the approach may be considered successful. Changes in single resistance components were closely correlated with changes in AUDPC: improvements in RC₁, RC₃ or RC₄ reduced AUDPC by the same, over the whole range of variation in infection frequency, affected leaf area, and infectiousness; on the contrary, little improvements in RC₂ were more effective than stronger ones. When components acted simultaneously, each of them reduced disease progress in proportion to its magnitude; when all components were improved by the same amount, they had about the same effectiveness in slowing the epidemic. Changing more components simultaneously reduced the disease development slightly more than additively. Advantages for plant breeders in improving their selection strategies are outlined.     

大豆品种早熟18抗疫霉根腐病基因的SSR分子标记

大豆品种早熟18是抗疫霉根腐病的有效抗源。本研究鉴定和分子标记早熟18的抗疫霉根腐病基因,以期为该品种的有效利用及分子辅助育种奠定基础。以感病大豆品种Williams与早熟18杂交建立分离群体。抗性遗传分析表明,早熟18对大豆疫霉菌抗性由1个显性单基因控制,该基因被定名为RpsZS18。SSR标记连锁分析表明,RpsZS18位于大豆分子遗传连锁群D1b上的SSR标记Sat_069和Sat_183之间,与这两个标记的遗传距离分别为10．0cM和8．3cM。RpsZS18是D1b连锁群上鉴定的第一个抗疫霉根腐病基因。     

用SSR标记分析抗疫霉根腐病大豆品种(系)的遗传多样性

利用50对SSR引物对抗疫霉根腐病大豆品种(系)进行遗传多样性分析。在166份品种(系)中,50个SSR座位共产生265个等位变异,平均每个座位5.3个。采用NTSYS-pc2.10计算品种(系)间遗传相似系数,平均相似系数为0.3124,表明抗疫霉根腐病大豆品种(系)间的遗传差异较大。用UPMGA进行聚类分析,166个品种(系)在相似系数为0.33时被聚为6类,地理来源相同的品种(系)大多聚类在一起。一些具有相同或相近抗病反应型的品种(系)被聚类在同一个类群中,表明这些抗病品种(系)的遗传关系较近,应有选择地利用。W illiam s和C lark抗疫霉根腐病近等基因系构成明显不同于中国大豆的基因源,可以用于拓宽我国大豆品种的遗传基础。