Cap G B, Roberts P A
Department of Nematology, University of California, Riverside 92521.
Electrophoresis. 1992 May;13(5):295-9. doi: 10.1002/elps.1150130161.
A relatively rapid and highly sensitive miniaturized polyacrylamide gel electrophoresis technique is described for the analysis of certain isozymes from single cotyledons of tomato seedlings and from single females of the root-know nematode (Meloidogyne spp.). Homogenates from single tomato cotyledons (7, 14, 21, and 28 days old) were electrophoresed and stained for acid phosphatase 1 (Aps 1) activity. Cotyledons from plants of all the above age groups showed good Aps 1 activity. Nondestructive screening for tomato Aps 1 is therefore feasible, using very small samples, from as young as 7-day-old tomato seedlings. This could be of important use in expediting root-knot nematode resistance (based on the Aps 1-linked resistance gene Mi) screening for breeding programs, or F1 testing for seed production purposes. In addition, the mini-polyacrylamide gel electrophoresis technique was useful for determination of the Aps 1 allelic contribution to the total enzyme activity. The system was also used to detect malate dehydrogenase and esterase isozyme activity from single adult females of the four common root-knot nematodes, Meloidogyne arenaria, M. hapla, M. incognita, and M. javanica, with equally good results, enabling species discrimination.
本文描述了一种相对快速且高度灵敏的小型聚丙烯酰胺凝胶电泳技术，用于分析番茄幼苗单片子叶和根结线虫（Meloidogyne spp.）单头雌虫中的某些同工酶。对不同日龄（7、14、21和28天）的番茄单片子叶匀浆进行电泳，并对酸性磷酸酶1（Aps 1）活性进行染色。上述所有年龄组植株的子叶均显示出良好的Aps 1活性。因此，使用非常小的样本，即便是7日龄的番茄幼苗，对番茄Aps 1进行无损筛选也是可行的。这对于加快根结线虫抗性（基于与Aps 1连锁的抗性基因Mi）筛选以用于育种计划，或用于种子生产目的的F1检测可能具有重要用途。此外，小型聚丙烯酰胺凝胶电泳技术有助于确定Aps 1等位基因对总酶活性的贡献。该系统还用于检测四种常见根结线虫，即南方根结线虫、北方根结线虫、爪哇根结线虫和花生根结线虫单头雌虫中的苹果酸脱氢酶和酯酶同工酶活性，结果同样良好，能够区分不同物种。
