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生化标记辅助筛选抗枯萎病的大蕉(Musa paradisiaca (L.) cv. puttabale)微繁殖克隆。

Biochemical markers assisted screening of Fusarium wilt resistant Musa paradisiaca (L.) cv. puttabale micropropagated clones.

作者信息

Krishna V, Kumar K Girish, Pradeepa K, Kumar S R Santosh, Kumar R Shashi

机构信息

P.G. Department of Studies and Research in Biotechnology and Bioinformatics, Kuvempu University, Shankaraghatta 577 451, India.

出版信息

Indian J Exp Biol. 2013 Jul;51(7):531-42.

PMID:23898552
Abstract

An efficient protocol was standardized for screening of panama wilt resistant Musa paradisiaca cv. Puttabale clones, an endemic cultivar of Karnataka, India. The synergistic effect of 6-benzyleaminopurine (2 to 6 mg/L) and thidiazuron (0.1 to 0.5 mg/L) on MS medium provoked multiple shoot induction from the excised meristem. An average of 30.10 +/- 5.95 shoots was produced per propagule at 4 mg/L 6-benzyleaminopurine and 0.3 mg/L thidiazuron concentrations. Elongation of shoots observed on 5 mg/L BAP augmented medium with a mean length of 8.38 +/- 0.30 shoots per propagule. For screening of disease resistant clones, multiple shoot buds were mutated with 0.4% ethyl-methane-sulfonate and cultured on MS medium supplemented with Fusarium oxysporum f. sp. cubense (FOC) culture filtrate (5-15%). Two month old co-cultivated secondary hardened plants were used for screening of disease resistance against FOC by the determination of biochemical markers such as total phenol, phenylalanine ammonia lyase, oxidative enzymes like peroxidase, polyphenol oxidase, catalase and PR-proteins like chitinase, beta-1-3 glucanase activities. The mutated clones of M. paradisiaca cv. Puttabale cultured on FOC culture filtrate showed significant increase in the levels of biochemical markers as an indicative of acquiring disease resistant characteristics to FOC wilt.

摘要

针对印度卡纳塔克邦的地方品种、抗巴拿马枯萎病的天堂蕉品种Puttabale克隆体,制定了一种高效的筛选方案。在MS培养基上,6-苄基腺嘌呤(2至6毫克/升)和噻苯隆(0.1至0.5毫克/升)的协同作用促使从离体分生组织诱导出多个芽。在6-苄基腺嘌呤浓度为4毫克/升和噻苯隆浓度为0.3毫克/升时,每个繁殖体平均产生30.10±5.95个芽。在添加5毫克/升BAP的培养基上观察到芽的伸长,每个繁殖体的平均长度为8.38±0.30厘米。为了筛选抗病克隆体,用0.4%的甲基磺酸乙酯对多个芽进行诱变,并在添加了尖孢镰刀菌古巴专化型(FOC)培养滤液(5-15%)的MS培养基上培养。通过测定总酚、苯丙氨酸解氨酶、过氧化物酶、多酚氧化酶、过氧化氢酶等氧化酶以及几丁质酶、β-1,3-葡聚糖酶等病程相关蛋白(PR蛋白)等生化标记物,对两个月大的共培养二次硬化植株进行抗FOC病筛选。在FOC培养滤液上培养的天堂蕉品种Puttabale的诱变克隆体,其生化标记物水平显著增加,表明获得了对FOC枯萎病的抗病特性。

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