【作者】 许艳丽；

【导师】 陈伊里；

【作者基本信息】 东北农业大学 ， 作物遗传与育种， 2004， 博士

【摘要】 大豆胞囊线虫病(Heterodera glycines Ichnohe)（SCN）是连作大豆主要病害和产生障碍的主要因素，大豆胞囊线虫病与连作大豆关系一直为人们所关注，本研究在国内首次采用长期定位田间试验方法，对北方典型的代表土类——黑土区大豆长期连作、豆麦豆迎茬、豆米豆迎茬、麦米豆轮作等主要轮作系统进行土壤环境对大豆胞囊线虫影响的研究，探讨不同土壤环境种植不同作物和不同大豆品种的根系分泌物和种植后土壤淋溶液与大豆胞囊线虫卵孵化的关系，并以在大豆胞囊线虫老病区育成的我国抗线品种为材料，对其抗病机制进行探讨。 描述了大豆长期连作、豆麦豆迎茬、豆米豆迎茬、麦米豆轮作等主要轮作方式下，土壤中大豆胞囊线群体变化情况。提出了大豆连作条件下土壤中胞囊多于其它茬口，豆米豆迎茬和豆麦豆迎茬次之，麦米豆轮作茬最少；但随着大豆连作年限的增加，连作区土壤中胞囊有逐步减少的趋势，认为可能是有抑制性土壤存在。 二十世纪八十年代以来我国相继育成了一些抗大豆胞囊线虫病品种，如在我国大豆主产区黑龙江省推广应用的抗线 1 号、2 号、3 号、4 号和 5 号等，但这些品种对大豆胞囊线虫抗性机制目前尚不清楚。本研究选用丰产性状好的抗线 4 号和当地主栽品种黑农35 进行在田间多种不同茬口上种植后大豆胞囊线虫发育动态系统研究，探讨抗线品种对大豆胞囊线虫抗性机制。试验结果表明，在大豆连作 12 年、豆米豆和豆麦豆茬口上种植的抗线 4 号根内 2 龄幼虫数量高于同茬口的黑农 35，抗线 4 号根内 3 龄幼虫和 4 龄幼虫数量明显低于黑农 35，抗线 4 号根面雌虫数量和收获后土壤中胞囊数量也低于黑农 35，认为抗线 4 号对大豆胞囊线虫抗性机制不是抗侵入，而是抗发育。首次报道了我国自己培育的抗大豆胞囊线虫品种抗线 4 号的抗性机制。 不同茬口条件下获得的大豆、玉米、甜菜、小麦和亚麻根渗出物对大豆胞囊线虫卵孵化均有刺激作用，甜菜根渗出物对大豆胞囊线虫卵孵化刺激作用最大，小麦和亚麻的根渗出物对大豆胞囊线虫卵孵化率受种植茬口影响相对较甜菜、大豆和玉米小；豆麦米茬口导致大豆根渗出物对大豆胞囊线虫卵孵化率明显提高，米豆米迎茬使得玉米根渗出物对大豆胞囊线虫卵孵化率增加。 不同茬口条件下种植的 5 种作物只有大豆种植后获得的土壤淋溶液都对大豆胞囊线虫卵孵化具有明显的促进作用，小麦、亚麻和玉米在一些茬口条件下表现出对大豆胞囊线虫卵孵化有抑制作用。大豆、玉米和甜菜在休闲区较其它茬口的土壤淋溶液对大豆胞囊线虫卵孵化具有明显的促进作用。 抗病品种抗线 3 号在豆麦米轮作、米豆米迎茬和豆麦豆迎茬土壤种植后获得的根渗出物对大豆胞囊线虫卵孵化具有刺激作用，尤其在米豆米茬口条件下刺激作用更为明显，在连作12年大豆和休闲条件下抗线3号根渗出物对大豆胞囊线虫卵孵化具有一定的抑制作用。合丰 25 在连作 12 年大豆条件下对大豆胞囊线虫卵孵化却具有促进作用，而在豆麦米轮作、麦豆麦迎茬、米豆米迎茬和豆麦豆迎茬条件下均表现为对大豆胞囊线虫卵孵化的抑制作用。 XI<WP=15>东北农业大学农学博士学位论文 种植抗、感大豆品种后 6 个茬口的土壤淋溶液对大豆胞囊线虫卵孵化率高于不种植大豆的土壤淋溶液和蒸馏水，说明 6 个茬口的土壤淋溶液在根系分泌物影响下对大豆胞囊线虫卵孵化都具有促进作用。感病品种合丰 25 种植后 6 个茬口的土壤淋溶液对大豆胞囊线虫卵孵化率高于抗病品种抗线 3 号，因此推断，感病品种合丰 25 的根系分泌物对土壤淋溶液有一定的影响。 豆麦豆迎茬、大豆 12 年连作、豆麦米、小麦 12 年连作和玉米 12 年连作轮作这些大豆茬口及休闲区土壤在不种植任何作物情况下土壤淋溶液都对大豆胞囊线虫卵孵化具有一定的抑制作用。 盐碱土条件下，种植抗感大豆品种后土壤淋溶液对大豆胞囊线虫卵孵化都具有明显的促进作用，并且种植抗感大豆品种后卵孵化率高于不种大豆的土壤淋溶液；白浆土种植感病品种合丰 25 后土壤淋溶液对大豆胞囊线虫卵孵化具有促进作用。由此说明盐碱土和白浆土大豆连作将导致大豆胞囊线虫发生严重，尤其种植感病品种危害将加重。 用限制性内切酶 AvaI 酶切大豆胞囊线虫群体的 rDNA-ITS 的 PCR 扩增产物后，使中国大豆胞囊线虫群体和美洲的加拿大、阿根廷和美国大豆胞囊线虫群体的 rDNA-ITS的 PCR 扩增产物产生不同的 RFLP 片断。表明用限制性内切酶 AvaI 可揭示中国和美洲的加拿大、阿根廷和美国大豆胞囊线虫(H. glycines)群体种内的遗传多样性，首次采用rDNA-ITS-RFLP 技术区分中国和加拿大、阿根廷大豆胞囊线虫群体。 首次将 ISSR 分析技术用于植物寄生线虫遗传多样性研究。筛选的 20 个 ISSR 引物中有 8 个引物可扩增出大豆胞囊线虫较清晰的谱带，用相应的引物可分析大豆胞囊线虫不同近亲系遗传多样性；用 FISSR2、FISSR5 和 FISSR6 引物扩增可得到加拿大、阿根廷、美国和中国的大豆胞囊线虫不同群体多态性的谱带。此结果说明，应用 ISSR 分子标记方?更多还原

【Abstract】 Soybean cyst nematode（Heterodera glycilnes Ichnohe,SCN）is one of maindiseases of soybean continuous cropping and a main barrier factor of those. Therelationship between soybean cyst nematode and soybean continuous croppingwas attented by researcher. First time using long term site-specific fieldexperiment to study soil environment effects on soybean cyst nematode ofdifferent rotation systems in black soil, the aim was to search the relationshipbetween soybean root exudates and soil leachate of different rotation systems andsoybean cyst nematode egg hatch. The resistance mechanism study aboutresistance variety Kangxian 4 which was breeded in China to soybean cystnematode was conducted. The molecular diagnosis with rDNA-ITS-RFLP and ISSR were used toidentify SCN　 population isolated from China, America, Canada, and Argentina.The study also focused on SCN　 populations China　 (Helongjiang isolates) as amodel to test the relationship between a major factors, plant root exudates, and thenematode egg hatch in the soil. The results showed soybean cyst nematode populations variety of the soil insoybean continuous cropping(S… S) 8-12 years, soybean rotation with corn andwheat(WCS) and soybeanrotation annually with wheat(or corn, SWS or SCS).Thecysts of soybean cyst nematode of the soil in S… SS for 8-12 years were more thanother rotation systems, the cysts of soybean cyst nematode of the soil in SWS andSCS were less and the cysts of soybean cyst nematode of the soil in WCS wereleast. As the years of soybean continuous cropping increase, the cysts of soybeancyst nematode in the soil had a trend to decrease. Perhaps it was the reason thatthere was suppressive soil . XIII<WP=17>东北农业大学农学博士学位论文 Since the 80’s of 20 centuries some resistance varieties to SCN such asKangxian 1, Kangxian 2, Kangxian 3,Kangxian 4 and Kangxian 5, were breeded inChina that were released in Heilongjiang province .But now the resistancemechanism of theirs were not know yet. In order to understand resistancemechanism of resistance variety Kangxian 4 relationship between the developmentdynamic of SCN and different soybean varieties was studied. The results showedresistance mechanismof resistance variety Kangxian 4 was resistance invasion. Soil leachate extracted from different crops cultivation showed differenteffects on the nematode egg hatch. Eggs treated with soil extract from soybeancultivation had significantly higher hatch rate than the ones treated with that fromother crops such as corn, sugar beet, wheat, and flax. Soil leachate extracted fromdifferent crops pre-cultivation showed positive effect on the nematodes egg hatch,in which beet pre-cultivation showed the most. This result explained why yearlyrotation between soybean and other crops can reduce the nematode damage.However, the soil extract from multi years continues cropping of soybean showedsuppression on the nematode egg hatch. This indicated that the concentration ofthe putative factors plays a role of regulating the nematode egg hatch. The amplification of the rDNA-ITS region of SCN populations from China,America, Canada, and Argentina yielded a single PCR fragment of approximately1060bp(including primes) for each of 16 SCN populations. No PCR products wereobtained in the control lacking the DNA template. A total 25 scored fragmentswere obtained with 17 restriction enzymes and used for analysis. The AvaI,EcoRI, HinfI, AluI, HaeⅢ, ClaI, MboI, AccI and RsaI digestions each showedidentical restriction profiles for all populations .The AvaI digestions of 5 ChineseSCN populations yielded 3 restriction fragments ,but yielded 2 restrictionfragments for America, Canada, and Argentina. It was the first report of geneticdiversity of SCN of Chinese , Canada, and Argentina with rDNA-ITS-RFLP. The genetic diversity of SCN populations from China, America, Canada, andArgentina were tested by ISSR analysis .The sel更多还原