一种新的组培大豆种子的消毒方法

A New Method for Disinfecting Soybean Seeds in Tissue Culture

为获得一种较好的环境友好型组培大豆种子的消毒方法,本试验通过对三种不同消毒方式下大豆种子组培4 d的萌发数目和污染数目进行统计。研究结果表明:通过不同浓度和不同时间的过氧化氢消毒后,四个品种的萌发数目趋势一致。都是随着15%H_(2)O_(2)消毒时间的延长,种子萌发越多,在灭菌1 h时全部萌发,且达到最大值;随着时间的再次延长,萌发数目减少且受到抑制。浓度为30%H_(2)O_(2)随着灭菌时间的延长,大豆种子萌发抑制程度加深,当灭菌2 h后,将近20%左右的大豆种子受到抑制未萌发。此外用同样的品种进行了其他两种方式的消毒,研究结果发现随着0.1%生汞、5%次氯酸钠消毒时间的增加,污染的种子越来越少,但是萌发数目越来越受抑制,在此试验品种中均没有达到完全消毒的目的。综上所述,选取15%H_(2)O_(2)消毒1 h最为合适,能够保证大豆种子完全萌发且不污染。同时选取国内主栽的15个大豆品种进行试验结果的验证,最终获得了一种新的组培大豆种子的消毒方法。此方法是一种高效、省时、便捷的消毒方式,这为下一步大豆遗传转化试验奠定了基础。

In order to obtain a better environment-friendly disinfection method for tissue culture soybean seeds,the number of germination and contamination of soybean seeds in tissue culture for four days under three different disinfection methods were counted.The germination numbers of the four varieties showed the same trend after different concentration and time of hydrogen peroxide disinfection.With the extension of 15%H_(2)O_(2) disinfection time,the more seeds germinated,and all seeds germinated at 1 hour of sterilization,and reached the maximum value.With the extension of time again,the number of germination decreased and was inhibited.With the extension of sterilization time,the germination inhibition of soybean seeds was deepened.After 2 hours of sterilization,about 20%of soybean seeds were inhibited and did not germinate.In addition,two other methods of disinfection were carried out with the same variety,and the results showed that with the increase of the disinfection time of 0.1%mercury and 5%sodium hypochlorite,the contaminated seeds were less and less,but the germination number was more and more inhibited,and the purpose of complete disinfection was not achieved in this test variety.In summary,it is most appropriate to select 15%H_(2)O_(2) for disinfection for 1 hour,which can ensure complete germination of soybean seeds without contamination.At the same time,15 main soybean varieties in China were selected to verify the test results,and finally a new method of tissue culture soybean seed disinfection was obtained.This method is an efficient,timesaving and convenient disinfection method,which lays a foundation for the next step of soybean genetic transformation test.