陈卓 肖熙鸥 陈曙 李可 邹华芬 金辉

摘    要：青枯病是危害马铃薯产业的重要病害，选育与推广抗青枯病品种是防控青枯病最高效的手段。目前已有的人工接种和田间抗性鉴定的方法不能有效区分处于潜伏侵染与完全抵抗侵染的材料，而利用GFP标记的Ralstonia solanacearum能够有效解决上述问题。笔者通过电击转化法将广宿主载体pBBR1MCS2-Tac-EGFP导入青枯菌P2中，成功获得了能够稳定遗传且不影响其致病力并带有绿色荧光报告的青枯菌菌株P2-Tac-EGFP。通过对感抗材料进行接种，结果表明，P2-Tac-EGFP能够有效区分感抗材料，并且P2-Tac-EGFP在感病材料的根部和茎部大量分布，而在抗病材料的根部仅有少量分布。综上所述，利用GFP标记的R. solanacearum能够快速准确地鉴定马铃薯青枯病抗性。

关键词：马铃薯;Ralstonia solanacearum;GFP;抗性

中图分类号：S532 文献标志码：A 文章编号：1673-2871（2021）01-035-07

Identification of potato bacterial wilt resistance using GFP-labeled Ralstonia solanacearum

CHEN Zhuo1， XIAO Xiou2， CHEN Shu2， LI Ke2， ZOU Huafen2， JIN Hui2

（1. College of Agriculture， Heilongjiang Bayi Agricultural University， Daqing 163000， Heilongjiang， China; 2. Key Laboratory of Tropical Fruit Biology， Ministry of Agriculture and Rural Affairs/Key Laboratory for Post-harvest Physiology and Technology of Tropical Horticultural Products of Hainan Province/Institute of South Subtropical Crop Research Institute， Chinese Academy of Tropical Agricultural Sciences  Zhanjiang 524091， Guangdong， China）

Abstract： Bacterial wilt is an important disease that endangers the potato industry. The selection and promotion of bacterial wilt resistant varieties is the most effective means to prevent and control bacterial wilt. The selection of germplasm resistant to bacterial wilt is the basis for the selection of resistant varieties. The current methods of artificial inoculation and field resistance identification cannot effectively distinguish between latent infection and disease-resistant materials， and Ralstonia solanacearum labeled with GFP can effectively solve the above problems. In this study， the broad host vector pBBR1MCS2-Tac-EGFP was introduced into R. solanacearum P2 by the electric shock transformation method， and successfully obtained the R. solanacearum strain P2-Tac-EGFP with stable inheritance without affecting its pathogenicity. Through inoculation of resistant materials， the results show that P2-Tac-EGFP can effectively distinguish the resistant materials， and P2-Tac-EGFP is distributed in a large amount in the roots and stems of the disease-resistant materials， while a small amount is in the roots of the resistant materials.In summary， the use of GFP-labeled R. solanacearum can quickly and accurately identify potato bacterial wilt resistance.

Key words： Potato; Ralstonia solanacearum; GFP; Resistance

馬铃薯（Solanum tuberosum L.）是我国四大主要粮食作物之一，也是世界上重要的作物之一，在我国的栽培面积仅次于小麦、水稻和玉米[1-2]。由茄科雷尔（青枯菌）式菌（Ralstonia solanacearum）引起的青枯病是马铃薯生产中的主要病害之一，能造成马铃薯大量减产，严重时可减产80% [3-5]。茄科雷尔式菌是一种危害维管系统的病原体，侵入根木质部并扩散到茎组织，在那里产生过量的胞外多糖及多种胞外蛋白堵塞和破坏导管，使植物失水萎蔫[6]。目前，使用抗病品种是马铃薯生产上最为经济有效的抗青枯病方式，但目前的马铃薯种质资源中抗性资源稀少[7]，因此，筛选马铃薯抗青枯病种质资源对马铃薯抗病育种起着关键性作用。……