摘要
褐飞虱(Nilaparvata lugens St(?)l.)是水稻主要害虫之一，应用抗褐飞虱品种是防治褐飞虱为害的有效手段。寄主植物对病虫的侵害具有两种遗传背景不同的抗性，即质量抗性和数量抗性。生产实践和众多的研究结果均表明，寄主植物的质量抗性很容易遭到害虫新的生物型的克服，缺乏稳定性和持久性；相比之下，数量抗性却要稳定和持久得多。定位与克隆水稻抗褐飞虱数量抗性基因是育种工作和遗传研究的共同目标。
B5和B14是药用野生稻与栽培品种杂交得到的转育材料，对褐飞虱表现为高抗。通过对TN1/B5和TN1/B14的F_2群体抗性分析，观察到TN1／B14群体的抗性级别分布接近3：1，而TN1/B5群体更接近正态分布。褐飞虱生物型1对B5和B14的反应在放虫5小时内略有差别，而生物型2对他们的反应基本没有差别。
为了近一步分析B5的抗性遗传背景，我们应用RFLP和SSR标记构建了MH63/B5重组自交系的分子遗传图谱。该图谱包括来源于Cornell大学和日本水稻基因组计划的156个RFLP标记和53个SSR标记，覆盖了水稻12条染色体的1，478cM，标记间平均距离为7.3cM。除了在第7和第8染色体分别有2个和1个断点外其余染色体上标记分布较为均匀，适合用于数量性状的遗传分析。多来源的分子标记使该图谱可以与已发表的其他水稻遗传图谱进行比较，有利于将来对其他图谱中信息的利用。
结合MH63/B5分子标记连锁图，我们尝试了对数量性状的动态分析方法，
以获得影响性状的基因在各个时间段的净遗传效应。以MH63旧5重组自交系
群体为材料，对分粟数进行了连续考察，然后应用“条件QTL”分析法对各
时段由遗传差异引起的分桑净增量进行了探讨。用混合线性模型进行分析，定
位了3个影响分葵能力的座位和相应的上位效应座位。相比常规QTL分析而
言，条件QTL更能体现遗传效应的时效性，对于发育相关的性状和其他发展
型性状的分析更合理、更可靠。动态QTL分析同样适用于褐飞虱抗性的研究。
应用常规QTL分析共定位了4个褐飞虱抗性遗传座位分别位于第2、3、4、9
染色体;应用条件Q几方法在第10染色体还多定位了一个座位。结果表明第
3、4染色体座位同黄臻定位的BPh14、BPh万位置相同，并有较大遗传效应，
其余座位效应较小。条件QTL的结果还表明BPh14、BPh乃的表达方式有很
大区别，即h14只在开始阶段表现出对抗性的贡献，BPh万的作用时间贯穿实
验始终。上位效应在和飞虱抗性中的作用较小。
王晓兰、袁红雨博士应用抑制减法杂交(SSH)cDNA文库，分离得到了
受褐飞虱取食诱导的水稻基因表达序列标签(EST)。我们通过基因组数据库
同源查找和RFLP定位将这些EST整合到连锁图谱上。建立一张水稻的抗褐
飞虱分子功能图谱。比较这些EST与褐飞虱抗性基因和QTL及前人报道的其
他抗性相关基因，观察到抗性基因和虫害诱导EST的聚集分布现象。还有17
个EsT与褐飞虱抗性基因或QTL的位置发生重合或临近。如果经过进一步遗
传分析能证实这些EST与褐飞虱抗性的关系，它们将成为和飞虱抗性候选基
因。
Brown planthopper (Nilaparvata lugens Stal., abbrievated as BPH) is one of the most serious pests of rice. Quality and quantitative resistance genes have been used in rice breeding. Many reports suggested that quantitative trait loci (QTL) are responsible for the durability of BPH resistance of rice. Development of molecular markers facilitates mapping of QTL. Mapping and cloning the BPH resistance QTL will be propitious to the development of resistant rice variety and understanding of BPH-resistance mechanism in rice.
B5 and B14 derived their resistance for BPH from wild rices. Genetic analysis suggested the distribution of BPH resistant plants to BPH susceptible plants in the TN1/B5 F2 population is about 3: 1, while it is about normal distribution in TN1/B14 F2 population. Different interaction between two biotypes of BPH and the two rice lines were observed in the first 5 hours. There was little difference between the number of biotype 1 BPHs settled on B14 and susceptible check in free choice test. But much fewer BPHs settled on B5.
A genetic map of rice covering 1,478 cM in the Kosambi function has been constructed using 187 F7 plants from a MH63/B5 RIL population. One hundred and fifty six RFLP markers from the Cornell University and Rice Genome Research Program (JRGP) and 53 SSR markers were located on the map using Mapmaker 3.0 with a LOD threshold 3.0. Except 3 gaps on chromosome 7 and 8, the markers distributed evenly on the map, and they are in good agreement with the
orders in the published maps of the Cornell University and JRGP. It is providing a cross-reference to the two sets of high-density linkage maps, and suitable for QTL analysis.
Dynamic analysis of QTLs for tillering ability based on the MH63/B5 RIL population was performed. Sequential records of tiller numbers of the RILs were analyzed by "Conditional mapping" approach, which was developed by Dr. Zhu for QTL detection of developmental trait, and traditional mapping. Three conditional QTLs and their net genetic effect were revealed. They showed higher statistic power than the QTLs detected in traditional mapping.
The interaction between BPH and rice is a time depended trait too. Damage scores of the parents and RILs were evaluated every five days from 12th day after infestation by the insects. Six QTLs contributing to BPH resistance were detected and mapped by using either conditional or traditional mapping. Five BPH resistant QTLs are derived from B5, and one was derived from MH63. The contributions to BPH resistance of the QTLs displayed different dynamic curves, and most of them were active in early stage of BPH infestation, but the effect of BphlS lasted to the end of the test (27 days after BPH infestation). The results are valuable to development of resistant rice variety and understanding the genetics of BPH-resistance in rice.
Dr. Wang and Yuan isolated Expressed Sequence Tags (ESTs) differentially regulated by BPH feeding by using techniques of the suppression subtractive hybridization (SSH) by. These ESTs were assigned to chromosomes based on RFLP mapping and rice genome database search. The distribution of the ESTs shows some level of clustering. Position coincidence of the ESTs and the BPH resistance genes/QTLs were observed. The feasibility of using differentially induced ESTs for candidate approach is discussed.
引文
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