Establishment of an artificial inoculation system for the efficient induction of rice bakanae disease

Yu-xin Yan , Xiao-yan Zhang , Yuan-yuan Tan , Jian-zhong Huang , Ljupcho Jankuloski , Qing-yao Shu
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引用次数: 1

Abstract

Bakanae disease, mainly caused by Fusarium fujikuroi, can lead to yield losses ranging from 10 to 20%. The lack of systematic researches on efficient propagation of pathogenic F. fujikuroi, artificial inoculation technology and the pathogenic conditions presents a critical technological gap that has severely hampered research on rice resistance to bakanae disease. This study first explored the sporulation conditions of F. fujikuroi in mung bean liquid medium, and determined that F. fujikuroi cultured in 40 ​g/L mung bean liquid medium for 2–6 days could efficiently produce pathogenic spores in large quantity. On this basis, we further establish an artificial inoculation system to induce bakanae disease. The main steps and parameters include: Soak rice seeds in a F. fujikuroi spore solution with a concentration of 1 ​× ​106 conidia/mL for 2 days; let imbibed seeds germinate, and choose germinating seeds with buds of a grain length and cultivate them on half strength MS medium (1/2 MS, pH 5.8–6.8) for about a week. The characteristic bakanae symptoms of excessively elongated seedlings could thus be induced. Furthermore, we found that the bakanae disease symptoms became more profound when a small amount of PDA (0.5%) was added to 1/2 MS medium. Hence our study established a technical system for the artificial induction of bakanae disease involving the use of mung bean medium for rapid propagation of F. fujikuroi spores, seed inoculation and cultivation of seedlings in 1/2 MS medium added with 0.5% PDA. This system will enable quick assessment of bakanae disease resistance and thus will be useful for rice breeding.

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建立有效诱导水稻白僵病的人工接种体系
Bakanae病主要由fujikuroi镰刀菌引起,可导致产量损失10%至20%。由于缺乏对病原菌的高效繁殖、人工接种技术和致病条件等方面的系统研究,造成了严重阻碍水稻抗bakanae病研究的关键技术空白。本研究首先探索了fujikuroi在绿豆液培养基中的产孢条件,确定了fujikuroi在40 g/L绿豆液培养基中培养2-6天,可以高效地产生大量致病孢子。在此基础上,进一步建立了诱导白卡菌病的人工接种体系。主要步骤和参数包括:水稻种子在浓度为1 × 106分生孢子/mL的fujikuroi孢子液中浸泡2天;让吸收的种子发芽,选择发芽种子,芽粒长度为1粒,在半强度MS培养基(1/ 2ms, pH值5.8-6.8)上培养一周左右。因此,可以诱导过长幼苗的特征性bakanae症状。此外,我们发现当在1/2 MS培养基中添加少量(0.5%)PDA时,bakanae疾病症状变得更加深刻。因此,本研究建立了一套利用绿豆培养基快速繁殖fujikuroi孢子,在添加0.5% PDA的1/2 MS培养基中接种种子,培养幼苗的人工诱导bakanae病的技术体系。该系统将能够快速评估bakanae的抗病性,从而对水稻育种有用。
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