Behura S K, Nair S, Sahu S C, Mohan M
International Centre for Genetic Engineering and Biotechnology, New Delhi, India.
Mol Gen Genet. 2000 Mar;263(2):328-34. doi: 10.1007/s004380051175.
In an attempt to identify a specific marker for biotype 2 of the Asian rice gall midge (Orseolia oryzae, Wood-Mason), we used AFLP (amplified fragment length polymorphism) fingerprinting. We identified an AFLP marker that is specifically amplified in biotypes 1, 2 and 5 of the rice gall midge, but not in biotype 4. Biotypes 1, 2 and 5 are avirulent to hosts bearing the Gm2 resistance gene (found in rice variety Phalguna), whereas biotype 4 is virulent to Gm2. Based on the sequence of this AFLP marker, SCAR (sequence characterized amplified region) primers were designed and used in combination with previously developed SCAR primers to distinguish effectively all five biotypes in a multiplex PCR-based assay. The inheritance pattern of this marker in the progenies of inter-biotype crosses between biotypes 1, 2 and 4 shows that the marker can be amplified by PCR from all F1 females, irrespective of the biotype status of their parents. However, the marker is present only in those male progenies whose mother was of a Gm2 avirulent biotype. The specific amplification of this marker in the avirulent biotypes and its pattern of inheritance show that avirulence with respect to carriers of the Gm2 gene in rice gall midge is sex-linked.
为了鉴定亚洲稻瘿蚊（稻瘿蚊，伍德 - 梅森）生物型2的特定标记，我们使用了扩增片段长度多态性（AFLP）指纹图谱技术。我们鉴定出一个AFLP标记，该标记在稻瘿蚊的生物型1、2和5中特异性扩增，但在生物型4中不扩增。生物型1、2和5对携带Gm2抗性基因（在水稻品种Phalguna中发现）的宿主无毒，而生物型4对Gm2有毒。基于该AFLP标记的序列，设计了序列特异性扩增区域（SCAR）引物，并与先前开发的SCAR引物结合使用，以在基于多重PCR的检测中有效区分所有五种生物型。该标记在生物型1、2和4之间的生物型间杂交后代中的遗传模式表明，无论其亲本的生物型状态如何，该标记都可以从所有F1雌性个体中通过PCR扩增出来。然而，该标记仅存在于其母亲为Gm2无毒生物型的雄性后代中。该标记在无毒生物型中的特异性扩增及其遗传模式表明，稻瘿蚊对水稻中Gm2基因携带者的无毒力是性连锁的。
