柑橘倍性检测方法的优化研究

Optimization study of citrus ploidy detection method

为了提高柑橘倍性检测效率以及降低倍性检测费用,进行了试剂用量、预处理方法以及检测液补水对柑橘倍性检测效果的试验。结果表明:25μL预混液处理后加入灭菌去离子水能正确检测出柑橘倍性;补水至混合液总体积在1000~1400μL范围内出峰效果较好,过多或过少均不利于检测出峰;同时将细胞核提取缓冲液和染色缓冲液按比例制成预混液,并采用解剖针粗头碾压的方式进行样品处理,试验效率比常规处理方法提高近2倍。综上,得到的优化方法能够大大降低柑橘倍性检测费用,实用方便,操作简单,在提高倍性检测效率的同时亦能提高试验操作的安全性,对柑橘品种改良以及倍性品种选育工作具有十分重要的意义。

In order to improve the efficiency of citrus ploidy detection and reduce the cost of ploidy detection,the effect of reagent dosage,pretreatment method and water supplementation of detection solution on the detection of ploidy of citrus was carried out.The results showed that citrus ploidy could be correctly detected by 25μL of master mix and then deionized water.The best effect of peak processing is that the amount of deionized water to the mixture in the range of 1,000~1,400μL,and too much or too little is not good for correct results.The processing efficiency is nearly 2 times higher than that of conventional methods,when the nuclei extraction buffer and staining buffer are proportionally made into a master mix,and the sample is pressed with the thick end of the dissecting needle.In summary,the optimization method obtained in this experiment can greatly reduce the cost of citrus ploidy detection,is practical and convenient,simple to operate,improves the efficiency of ploidy detection and improves the safety of test operation,which is of great significance for the improvement of citrus varieties and the breeding of ploidy varieties.