Gengenbach B G, Green C E, Donovan C M
Department of Agronomy and Plant Genetics, University of Minnesota, St. Paul, Minnesota 55108.
Proc Natl Acad Sci U S A. 1977 Nov;74(11):5113-7. doi: 10.1073/pnas.74.11.5113.
Texas male-sterile cytoplasm (cms-T) maize is susceptible to Helminthosporium maydis race T and its pathotoxin, whereas nonsterile cytoplasm maize is resistant. Callus cultures initiated from immature embryos of a cms-T genotype, BC(1)A188(T), were susceptible to the toxin and were capable of plant regeneration. Toxin-resistant cell lines were selected by a sublethal enrichment procedure in which cms-T callus was grown for several selection cycles (subculture transfers) in the presence of progressively higher concentrations of toxin. Periodically during the selection process, plants were regenerated from the cms-T cultures to determine their susceptibility or resistance to the toxin. Plants regenerated after four cycles of selection were male-sterile and toxin-susceptible as shown by leaf bioassays. All plants regenerated from cell lines isolated from the fifth selection cycle onward, however, were toxin-resistant and 52 of 65 were fully male-fertile. The remaining 13 "male-sterile" resistant plants did not shed pollen and did not resemble cms-T plants in tassel morphology. Some "male-sterile" plants produced anthers containing a small amount of starch-filled pollen, suggesting that the sterility of these 13 plants was not the result of the cms-T trait. Leaf bioassays on F(1) progeny from regenerated resistant plants indicated that resistance to the toxin was inherited only through the female. The male-fertility trait also was inherited only through the female. After inoculation with H. maydis race T spores, leaf lesion size for progeny from regenerated resistant plants coincided with their reaction to the toxin. This result indicated that plant resistance to the pathogen was closely correlated with the toxin resistance obtained through cell culture selection.
德克萨斯雄性不育细胞质(cms-T)玉米对玉米长蠕孢菌T小种及其致病毒素敏感,而非不育细胞质玉米则具有抗性。从cms-T基因型BC(1)A188(T)的未成熟胚诱导产生的愈伤组织培养物对该毒素敏感,且能够再生植株。通过亚致死富集程序筛选出毒素抗性细胞系,在此过程中,cms-T愈伤组织在逐渐升高的毒素浓度下进行几个选择周期(继代转移)的培养。在选择过程中定期从cms-T培养物再生植株,以确定它们对毒素的敏感性或抗性。四轮选择后再生的植株经叶片生物测定表明为雄性不育且对毒素敏感。然而,从第五轮选择周期开始分离得到的细胞系再生的所有植株均对毒素具有抗性,65株中有52株完全可育。其余13株“雄性不育”抗性植株不散落花粉,其雄穗形态也与cms-T植株不同。一些“雄性不育”植株产生的花药含有少量充满淀粉的花粉,这表明这13株植株的不育并非cms-T性状所致。对再生抗性植株的F(1)后代进行叶片生物测定表明,对毒素的抗性仅通过母本遗传。雄性可育性状也仅通过母本遗传。用玉米长蠕孢菌T小种的孢子接种后,再生抗性植株后代的叶片病斑大小与其对毒素的反应一致。这一结果表明,植株对病原菌的抗性与通过细胞培养选择获得的毒素抗性密切相关。