【作者】 王兴春；

【导师】 杨长登；

【作者基本信息】 中国农业科学院 ， 作物遗传育种， 2003， 硕士

【摘要】 稻瘟病、白叶枯病、白背飞虱和褐飞虱是水稻生产上的主要病虫害，培育和推广抗性品种是持续控制病虫害流行最经济、安全、有效的措施。然而，目前生产上推广的多为单抗品种，仅能抵抗某种病虫的危害，并且其抗性是由主效基因控制的，一般只能维持3～5年时间。聚合多个抗性基因或利用微效多基因是延长抗病虫品种抗性周期的最有效办法；构建对多种病虫害有较好抗性的多抗材料，是未来抗性育种的发展方向。 本研究将分子标记辅助选择和花药培养等现代生物技术与回交转育等传统技术有机结合起来，建立了快速转育和聚合水稻抗性基因的技术体系。并应用该体系，以五丰占2号为轮回亲本，IRBB5为非轮回亲本，经过一次回交，成功地将抗白叶枯病隐性主效基因xa—5快速转育五丰占2号中，获得了将白叶枯病抗性隐性主效基因与微效多基因聚合，且抗稻瘟病、白背飞虱和褐飞虱的材料。主要研究结果如下： 1．IRBB5携带有抗白叶枯病隐性主效基因xa-5，其与感病亲本蜀恢162所配组合的分离后代表现了一对隐性主基因的分离比：五丰占2号的白叶枯病抗性受微效多基因控制，基因效应分析表明，该性状符合加性-显性模型，以加性效应为主。 2．在五丰占2号2／IRBB5的B₁F₁群体中，白叶枯病抗性达到五丰占2号水平的植株数占68.3％，说明一次回交就可选育到白叶枯病抗性与五丰占2号水平相当又携带有xa-5基因的植株。 3．设计出一条新的与白叶枯病抗性基因xa-5紧密连锁的CAPS引物5′CCAGACACCACTGCACATTC3′，该引物GC含量适中，扩增效率更高。然而，对双亲扩增产物的测序结果表明，扩增产物序列同源性很高，达96％，不能设计出三条特异引物，进行三引物标记。因此，要简化xa-5分子标记程序还须另寻它法。 4．接种花药25560枚，出愈数为1536个，出愈率为6.0％；得到62株绿苗，绿苗率为0.24％。其中，一次成苗培养基、M₈培养基和SH培养基花培绿苗率分别为0.37％，0.21％和0.14％，表明一次成苗培养基更适合于本研究的籼稻花药培养。 5．花药培养和分子标记辅助选择等现代生物技术与传统回交育种技术的有机结合，大大加快了多抗材料构建的进程，从配组到获得聚合纯系只用了 2年时间。6．获得了3份聚合有白叶枯病抗性隐性基因xa－5和微效多基因，且抗稻瘟 病。白背飞虱和褐飞虱的多抗材料。其中WZ高抗白叶枯病，抗稻瘟病。 白背飞虱和褐飞虱。7．多抗品系 WZ的米质结合了双亲的优点，与五车占 2号相比，门项米质 指标中除了 5项相似外，其余 6项得到明显改善，即要自率山 22 ％下降 到 l％，要白度山 4．2％降低为 0．4％，透明度从 3级提高到］级，碱消值 从3．2提高到7．0，胶稠度山59mm提高到78mm，直链淀粉含量从14．7o 提高到 18．4％。说明经过一次回交后，可以剔除某些不良基因，从而选育 出品质较好的多抗品系。更多还原

【Abstract】 Blast, bacterial blight and planthopper are the important diseases or insect pests in rice production. Development of resistant varieties was considered to be the most effective, efficient and environment-friendly strategy controlling the diseases and pests. However, the resistant varieties in use could only resist to one or two diseases or pests and the resistance was conferred by single major gene. Most of them would be broken down after 3-5 year’s release. Therefore, development of multi-resistant varieties was the hot issue in resistance breeding. Establishment durable resistance was the key problem.In this study, the integrating technology of anther culture, MAS and backcross was developed and successfully used in transferring xa-5, a recessive gene for resistance to bacterial blight, from IRBB5 to Wu Feng Zhan 2(WFZ2). Gene pyramiding lines resistance to bacterial blight, blast and planthopper were obtained. The results were as follows:1. Segregation Population derived from IRBB5 with a recessive gene for resistance to bacterial blight and Shuhui 162 was inoculated with bacterial blight pathogen Zhel73. The inoculation demonstrated one major recessive gene segregation ratio. The bacterial blight resistance of WFZ2 was controlled by minor-polygene. The character conformed to additive-dominant model and was mainly controlled by additive effects.2. In the B1F1 population of WFZ2 and IRBB5, 68.3% plants got the bacterial blight resistance as recurrent parent WFZ2. It indicated that the plants with the same resistance level to WFZ2 and xa-5 gene could be obtained through one time backcross.3. A new revise primer, 5’ CCAGACACCACTGCACATTC3’ , was designed according to the sequence of P0574H01 in Gene Bank using Primer3. With a rational GC ratio, the new revise primer had a similar Tm to the forward one. The PCR amplification efficiency was greatly improved. Unfortunately, the sequence comparison of two parents indicated that they were too similar, about96%, to design tri-primer to amplify. To simplify the PCR procedure, other method should be taken.4. One thousand five hundred and thirty six calli and 62 green plantlets were obtained with 6.0% callus induction frequency and 0.24% green shoot regeneration frequency. The green shoot regeneration frequencies were 0.37% with PPA-based one-step medium, 0.21% with M8 medium and 0.14% with SH medium. It indicated that the PAA based one-step medium was most effective in the anther culture of indica genotypes.5. The integrating technology of anther culture, MAS and traditional backcross was efficient in multi-resistant breeding, only two years from the cross to the pure lines with multi-resistance.6. Three multi-resistant lines with xa-5 and minor-polygene for bacterial blight and resistance to rice blast and planthopper were obtained. Among of them, W2 showed high resistance to bacterial blight, resistance to blast and planthopper.7. Quality test of W2 showed that: among of the 11 quality standards, there were 5 similar to WFZ2 and 6 were improved, namely chalky grain from 22% to 1%, chalkiness from 4.2 to 0.4, translucency from 3 to 1, alkali spreading value from 3.2 to 7.0, gel consistency from 59mm to 78mm and amylose content from 14.5% to 18.4%. It indicated that some defective characters in WFZ2 could be sorted out through one time backcross and high quality line with multi-resistant genes could be obtained.更多还原