酶标仪在稻米表观直链淀粉含量测定及突变体筛选中的应用

探索了 96孔全自动酶标仪 (SpectraMax 1 90 ,Sunnyvale,USA)在稻米表观直链淀粉含量(AAC)测定中的应用效果。实验以IR2 4、Iri371、Z41 3、KMD1、秀水 1 1共 5个品种为材料 ,以农业部部颁标准为参照 ,样本处理后不作定容稀释而直接在 96孔酶标板上进行显色 ,测定OD值 ,根据标样获得的回归等式计算待测样本的AAC。结果表明 ,酶标法与国标法测得的精度无显著差异。利用这一体系 ,对 1 5 0 0余份协青早B的M3 (收自M2 单株 )种子进行了AAC突变体筛选及后代跟踪研究 ,获得了 1份中等AAC( 1 6%左右 )突变体ZXY1 6。该方法操作简便易行 ,据此可建立高通量稻米AAC测定体系

APPLICATION OF MICROPLATE SPECTROPHOTOMETER IN DETERMINATION OF APPARENT AMYLOSE CONTENT AND MUTANT SELECTION IN RICE

The application of automatic microplate spectrophotometer (SpectraMax 190, Sunnyvale, USA ) was investigated for measurement of apparent amylose content (AAC) of rice grains. By using five rice varieties (IR24, Iri371, Z413, KMD1 and Xiushui 11), a microplate spectrophotometer-facilitated AAC measurement system was established with similar accuracy to that of standard protocol (NY147-88). In this system, sample was treated as NY147-88, but directly stained in a microplate and the OD values of each sample were determined by a microplate spectrophotometer. A regression equation was established by using the AAC and OD value of standard samples, and the AAC of test samples were estimated by using the equation. The system is simple, accurate and high-throughput. By using this system, about 1500 progenies of M-3 seeds (harvested from individual M-2 plants) were screened for putative AAC mutants, from which one mutant line ZXY16 was developed with a middle AAC of about 16%.