【作者】 王月明；

【导师】 王冬梅；

【作者基本信息】 河北农业大学 ， 植物学， 2006， 硕士

【摘要】 大豆花叶病毒(Soybean Mosaic Virus,简称SMV)病是世界性大豆病害之一,也是我国各大豆产区普遍发生的重大病害之一,对大豆生产危害严重,造成产量损失,籽粒品质下降。SMV以种子带毒为初侵染源,蚜虫以非持久方式传毒,使SMV容易造成流行性危害。由于化学药剂不能有效地防治该病害,所以推广抗病品种是目前公认的最经济有效的病毒防治方法。但是不少地区反映,从外地引入的抗病品种往往不能解决本地区的大豆花叶病危害问题。这是由于各地区大豆花叶病毒的流行株系不同,因而对不同的大豆品种有不同的致病力,大豆品种对不同株系的抗病性反应也有很大的差异。因此,在选育抗病品种时,必须查明本地区的大豆花叶病毒的流行株系和株系组成,有的放矢地筛选抗原,这是一项经济有效的工作。 为调查河北省推广大豆品种对大豆花叶病毒的抗性情况,本研究对9份大豆品种,包括高蛋白品种:冀豆12号,冀豆7号;高油品种:冀黄13号,nf37,nf58:兼性品种:冀豆15号,鉴15;以及无腥大豆品种:五星1号,五星2号,均采用人工汁液摩擦法分别接种六个SMV株系,包括河北省的流行株系Sc-8,Sc-11和一个待定株系3238;黄淮地区的Sc-3;江苏的Sa:东北的N3进行抗性鉴定。鉴定结果表明:在供鉴的九份大豆品种中,高蛋白品种冀豆12号是一个较理想的抗病毒品种,它对试验中所用的6个病毒株系均表现1级反应型;2001年培育成功的无腥品种五星1号是一个优良的抗病毒品种,它对供试的6个株系中的5个表现0级反应型,对来自东北的病毒株系N3表现1级反应型;五星2号的抗病毒能力也较强。而高油品种的抗病毒能力都表现不太理想。由此认为五星1号、冀豆12号和五星2号是三个较理想的抗SMV品种,适合推广种植。 在此基础上,我们从中选出一对抗感组合,大豆品种冀豆7号对SMV株系N3为抗病组合,对SMV株系Sc-8为感病组合,并对其侵染机制进行了初步的研究。本试验运用电子显微镜技术初步研究了大豆花叶病毒毒株Sc-8和N3侵染冀豆7号后产生的一系列细胞病理学变化。接种病毒后于不同时间点采集冀豆7号叶片,经过化学固定、脱水和包埋后制作超薄切片,切片经染色后在透射电镜下观察。在抗病组合的叶肉细胞中,侵染早期(接种8-12h),叶绿体膨胀,叶绿体片层结构轻微零乱,核染色质发生凝集现象;接种后24h,叶绿体继续膨胀变形,线粒体结构清晰完整,核变形严重;侵染后期(接种后72h),细胞核近乎衰败,双层核膜已基本辨认不清,叶绿体结构基本解体。此时线粒体嵴突已发生退化,只有双层膜结构,内部出现空虚状态。细胞中的病毒粒子很少,也没发现风轮状内含体结构,充分体现了病毒增殖受到抑制的情况。在此过程中,叶绿体和细胞核是最早做出反应的细胞器,而线粒体是最后解体的细胞器。叶肉细胞在超微结构水平的变化显示出了植物细胞程序性死亡(program cell death,PCD)的某些形态特征。感病组合叶肉细胞超微结构变化比抗病组合晚了10多个小时,细胞器的变化与抗病组合相似,需要说明的一点是在所观察的整个互作过程中,核、叶绿体和线粒体的衰退是同步的,显示出了细胞被动死亡的更多还原

【Abstract】 Soybean mosaic virus (SMV) occurs in soybean worldwide and causes significant yield loss and seed quality reduction. The virus is seed-transmitted and also readily sap-and aphid-transmissible, so easily cause prevalence. Because SMV can’t be prevented by chemicals, many resistance cultivars have developed and used in soybean production. But , the cultivars indraught from other regions can not resistant to their own region’s virus in fact. It is because there are different SMV strains in different regions, and cultivar’s resistance extent are not the same;also, SMV’s infection ranges are not the same. Based on these reasons, when we choose the resistance cultivars, we must investigate the region’s prevalence SMV strains and then study resistance identification. This is an economic and efficiency work.To study our province’s soybean cultivar’s resistance, we choosed nine soybean cultivars mainly planted in Hebei province, including high protein content cultivars: Jidou 12, Jidou 7;high oil content cultivars: Jihuang 13, nf37, nf58;both high content of protein and oil cultivars: Jidoul5, Jian15;and cultivars without fishy smell: Wuxing 1, Wuxing 2, manually rub inoculation with six strains of SMV, including Sc-8, Sc-11,3238 separated and purification from Hebei province, Sc-3 from Huanghuai region, Sa from Jiangsu, N3 from Northeast. The results showed that: The high protein content cultivar Jidou 12 is an ideal resistance cultivar, which exhibits the first type of resistance level to six strains;Wuxing 1 breeded in 2001 exhibits the zero type of resistance level to five in six strains;and Wuxing 2 also a good resistant sources to SMV. But the high oil content cultivars all exhibit lower resistance to six strains. So we believed Wuxing 1, Jidou 12 and Wuxing 2 are three good resistant sources to SMV.We selected a virus resistant combination on the basis of identification above. We found that jidou 7 is sensitive to Sc-8 of SMV and resistant to N3 strain of SMV .Electron microscope technology was used to reveal the mechanism of soybean how to resist the virus infecting. In our research, a series of cytopathic effects were found in jidou 7 after infected by Sc-8 and N3 strans. After jidou 7 was inoculated with virus, the leaves were taken at different time to get the ultrathin sections observed under transmission electron microscope. The cytology changes after the virus infecting in incompatible combination are: at the early stage (inoculated 8 to 12 hours), the structure and shape of chloroplasts and nuclei had a little change;at the later stage (inoculated 72 hours), cell nucleus agglutinated and degradation, the membrane of nuclei and chloroplasts disappeared. Mitochondriadegradation only have two layer of membrane structure. There are only a few viruses observed in the cells , and the pinwheels were not found in cytoplasm. These results indicated that the virus have been restrained in incompatible combination. Duiring the course, chloroplasts and nuclei are the first changed organes, but mitochondria are the last degradated organ. These showed some characters of programmed cell death. In compatible combination the cytology symptoms changed 10 hours later than the incompatible combination . we found pinwheel inclusion in cytoplasm at 5 days after inoculated. Otherwise, the starches synthesized in all the time before cell breakdown. In the cytoplasm the number of mitochondria increased abnormaly, perhaps for the sake of energy anabolism for virus multiplication. In addition, we proved that virus RNA begin to transport with long distance at 12 hours after inoculation.更多还原