为改进甘薯组培苗的病毒检测工艺,以甘薯病毒指示植物巴西牵牛(Ipomoea setosa)种子在MS+GA3 2.0 mg/L培养基上萌发苗的茎尖为外殖体,在MS+IBA 0.2 mg/L的培养基上茎尖能够发育成完整的试管植株且能够以单叶节扦插通过腋芽再植株生长方式快速繁殖;在试管中将巴西牵牛试管苗和甘薯试管苗茎段进行试管微嫁接后在MS0培养基上培养,15天后,与带病毒的甘薯试管苗嫁接的巴西牵牛茎段所带的叶片均表现出黄化、花叶等阳性反应。与不带病毒的甘薯试管苗茎段嫁接的巴西牵牛试管苗茎段所带的叶片都正常生长;用甘薯试管苗的无菌研提汁液感染巴西牵牛试管苗茎段切口后在MS0培养基上培养,15天后,被带病毒甘薯试管苗研提汁液感染的巴西牵牛试管苗茎段所带的叶片都表现出黄化、花叶等阳性反应,继续培养叶片枯死。被不带病毒甘薯试管苗研提汁液侵染的巴西牵牛试管苗茎段所带的叶片都能正常生长;以上2种方法可以替代传统的“巴西牵牛种子苗网室嫁接检测法”检测甘薯组培苗是否带有病毒。具有灵敏度高、检测结果不受外界环境影响、检测成本低、培养条件可控、不受季节性限制等优点。
In order to improve the detection technologies of the viruses in test-tube plantlets of sweet potato, we have been rapid propagation the test-tube plantlets of ipomoea setosa in the medium “MS+IBA 0.2 mg/L” using the growing point on the stem of Ipomoea setosa that sprouting by the seeds in the medium “MS+GA3 2.0 mg/L”. Grafted the test-tube plantlets of sweet potato and the test-tube plantlets of Ipomoea setosa and cultured in the medium “MS0” in vitro, after 15 days. The leaves of the test-tube plantlets of Ipomoea setosa that grafting with the test-tube plantlets of sweet potato infected by the virus become yellowing or floral leaf. The test-tube plantlets of ipomoea setosa that grafting with the test-tube plantlets of sweet potato no virus infection can normal growth. Cultured the test-tube plantlets of Ipomoea setosa that infected by body fluid of the sterile test-tube plantlets of sweet potato through the wounds in stem in the medium “MS0” in vitro, after 15 days. The leaves of the test-tube plantlets of ipomoea setosa that infected by containing the virus body fluid of the sterile test-tube plantlets of sweet potato become yellowing or floral leaf then exsciccation. The test-tube plantlets of Ipomoea setosa that infected by no virus body fluid of the sterile test-tube plantlets of Sweet Potato can normal growth, both methods can displace the old methods that “Grafted the plantlets of Sweet Potato and the plantlets of ipomoea setosa in net room”. It has the advantages of high sensitivity, low cost, easily controlled culture conditions, not limited by season, not affected by the external environment, etc.
张希太 张彦波 肖磊 谢淑芹 .利用巴西牵牛试管苗检测甘薯组培苗病毒技术研究.农学学报 ,2013,3(12): 16-22