芦丁和IAA对绿豆黄化幼苗伸长的影响及对相关基因的RT-PCR

就芦丁(20~100μg/mL)、IAA(0.5~2.0μg/mL)及芦丁(40μg/mL)加IAA(0.5~2.0μg/mL)对绿豆黄化幼苗伸长的影响进行了研究.结果表明,芦丁对黑暗中生长的绿豆黄化幼苗下胚轴和胚根的伸长均有抑制作用,添加IAA促进了这种抑制作用.RT-PCR扩增产物的分析表明,绿豆黄化幼苗经芦丁和IAA分别处理或混合处理之后,其体内吲哚氨基酸水解酶基因、吲哚-3-甘油磷酸合成酶基因及查尔酮合成酶基因的表达在不同处理间表现出差异和多态性,芦丁处理后,其吲哚-3-甘油磷酸合成酶基因不表达.不同的RNA提取程序的比较认为,异硫氰酸胍有助于绿豆黄化幼苗RNA的提取.RT-PCR反应中不同退火温度的比较显示,较高的退火温度(60℃)有助于特异性扩增.

Effects of rutin and IAA on elongation of etiolated mung bean (Vigna radiata ) seedlings and RT-PCR diversity of related genes

Effects of rutin (20- 100 μg/mL), indoleacetic acid (IAA 0.5 - 2.0 μg/mL) and rutin (40 μg/mL)plus IAA (0.5- 2.0 μg/mL) on elongation of etiolated mung bean seedlings were observed. The experimental results showed that elongation of the seedlings was inhibited by rutin and the inhibiting action was enhanced by IAA. After the seedlings were treated by rutin, IAA and rutin plus IAA, the expressions indicated by RT-PCR of genes encoding indole-amino-acid hydrolase, indole-3-glycerol phosphate synthase and chalcone synthetase,respectively, displayed differences among different above treatments. The IGPS gene was not expressed in the etiolated seedlings treated with 40 μg/mL rutin. Comparison of different RNA extraction procedures indicated that guanidinium isothiocyanate was important for the RNA extraction. Differences of RT-PCR amplificates at different annealing temperature showed that higher annealing temperature (60 ℃ ) was helpful to specific amplification.