生物技术通报 ›› 2023, Vol. 39 ›› Issue (4): 221-226.doi: 10.13560/j.cnki.biotech.bull.1985.2022-0994
李月1(), 余婉贤1, 李宁2, 姚明华2, 李峰1(), 邓颖天1()
收稿日期:
2022-08-10
出版日期:
2023-04-26
发布日期:
2023-05-16
通讯作者:
李峰,博士,教授,研究方向:小分子RNA在植物病原互作中的功能;E-mail: chdlifeng@mail.hzau.edu.cn;作者简介:
李月,女,硕士研究生,研究方向:辣椒炭疽病;E-mail: 2465461425@qq.com余婉贤为共同第一作者
基金资助:
LI Yue1(), YU Wan-xian1, LI Ning2, YAO Ming-hua2, LI Feng1(), DENG Ying-tian1()
Received:
2022-08-10
Published:
2023-04-26
Online:
2023-05-16
摘要:
炭疽病是辣椒生产上所面临最严重的病害之一,迫切需要可靠的方法在辣椒发育早期筛选抗病资源。炭疽病菌株CD的分生孢子采用两种接种方法,对辣椒‘ST-8’不同苗龄材料进行接种,通过对发病率、叶片卷曲率以及病斑大小的比较分析最佳的接种方法。结果表明,使用浓度为1×106个/mL的孢子悬浮液进行针刺点接法效果最佳,有望应用于大规模高通量辣椒抗炭疽病种质筛选。
李月, 余婉贤, 李宁, 姚明华, 李峰, 邓颖天. 辣椒苗期炭疽菌接种方法[J]. 生物技术通报, 2023, 39(4): 221-226.
LI Yue, YU Wan-xian, LI Ning, YAO Ming-hua, LI Feng, DENG Ying-tian. Inoculation Method for Colletotrichum in Pepper(Capsicum annuum)Seedlings[J]. Biotechnology Bulletin, 2023, 39(4): 221-226.
图1 针刺接种法示意图 A: 接种过程中(箭头:针刺点); B: 接种后(箭头:接种点)
Fig. 1 Needle point method A: During inoculation(arrowheads: needle point position). B: After inoculation(arrowheads: inoculated position)
图2 辣椒苗4个时期 A:子叶期; B:1-2片真叶期;C:3-4片真叶期;D:5-10片真叶期
Fig. 2 Four different stages of pepper seedlings A: Cotyledonary stage; B: 1st - 2nd leave stage; C: 3rd - 4th leave stage; D: 5th - 10th leave stage
图3 利用ImageJ批量测量病斑大小 A:设定照片比例尺; B:选中并添加病斑区域;C:测量病斑面积
Fig. 3 Batch measurement of lesion area by using ImageJ A: Set up photo scale. B: Choose lesion area. C: Measure chosen areas
孢子悬 浮液浓度 Resuspend spore concentration (cells·mL-1) | 子叶期 Cotyledon | 1-2片真叶期 1st - 2nd leave | 3-4片真叶期 3rd - 4th leave | 5-10片真叶期 5th - 10thleave | 果实 Fruits | |||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
针刺法Needle point | 针刺法 Needle point | 注射法 Infiltration | 针刺法 Needle point | 针刺法 Needle point | 针刺法 Needle point | |||||||
感病率 Infe rate/% | 叶卷率 Curlrate/% | 感病率 Infe rate/% | 叶卷率 Curl rate/% | 感病率 Infe rate/% | 叶卷率 Curl rate/% | 感病率 Infe rate/% | 叶卷率 Curl rate/% | 感病率 Infe rate/% | 叶卷率 Curl rate/% | 感病率 Infe rate/% | ||
1×103 | 54.17 | 67.86 | 10.81 | 41.89 | 85.71 | 14.29 | 6.06 | 0 | 6.90 | 0 | 38.89 | |
1×104 | 100.00 | 75.00 | 60.00 | 36.00 | 100.00 | 5.00 | 61.11 | 0 | 20.83 | 0 | / | |
1×105 | 100.00 | 96.43 | 100.00 | 78.57 | 100.00 | 25.00 | 80.00 | 0 | 54.55 | 0 | 100.00 | |
1×106 | 100.00 | 100.00 | 100.00 | 80.65 | 100.00 | 41.67 | 84.38 | 0 | 72.00 | 0 | / | |
1×107 | 100.00 | 100.00 | 100.00 | 91.94 | 100.00 | 62.50 | 90.00 | 2.50 | 87.50 | 0 | 100.00 |
表1 辣椒不同苗期接种不同浓度炭疽菌后感病率与叶片卷曲率统计
Table 1 Statistical analysis of plant infection and leave curl rates of pepper seedlings in different stages under different Colletotrichum spore concentration
孢子悬 浮液浓度 Resuspend spore concentration (cells·mL-1) | 子叶期 Cotyledon | 1-2片真叶期 1st - 2nd leave | 3-4片真叶期 3rd - 4th leave | 5-10片真叶期 5th - 10thleave | 果实 Fruits | |||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
针刺法Needle point | 针刺法 Needle point | 注射法 Infiltration | 针刺法 Needle point | 针刺法 Needle point | 针刺法 Needle point | |||||||
感病率 Infe rate/% | 叶卷率 Curlrate/% | 感病率 Infe rate/% | 叶卷率 Curl rate/% | 感病率 Infe rate/% | 叶卷率 Curl rate/% | 感病率 Infe rate/% | 叶卷率 Curl rate/% | 感病率 Infe rate/% | 叶卷率 Curl rate/% | 感病率 Infe rate/% | ||
1×103 | 54.17 | 67.86 | 10.81 | 41.89 | 85.71 | 14.29 | 6.06 | 0 | 6.90 | 0 | 38.89 | |
1×104 | 100.00 | 75.00 | 60.00 | 36.00 | 100.00 | 5.00 | 61.11 | 0 | 20.83 | 0 | / | |
1×105 | 100.00 | 96.43 | 100.00 | 78.57 | 100.00 | 25.00 | 80.00 | 0 | 54.55 | 0 | 100.00 | |
1×106 | 100.00 | 100.00 | 100.00 | 80.65 | 100.00 | 41.67 | 84.38 | 0 | 72.00 | 0 | / | |
1×107 | 100.00 | 100.00 | 100.00 | 91.94 | 100.00 | 62.50 | 90.00 | 2.50 | 87.50 | 0 | 100.00 |
图4 比较注射法与针刺法接种炭疽菌于辣椒苗期叶片 A-F:注射法;G-L:针刺法;标尺=10 mm
Fig. 4 Comparing inoculated pepper seedling leaves by using infiltration and needle point method A-F: Infiltration method. G-L: Needle point method. Bar = 10 mm
图5 利用针刺法对不同时期辣椒苗接种不同浓度炭疽菌后叶片发病对比
Fig. 5 Colletotrichum inoculated leaves by using needle point method during different developmental stages of pepper seedlings at different Colletotrichum spore concentration
图6 利用针刺法接种炭疽菌于辣椒果实与苗期对比 A-D:利用针刺法对辣椒果实接种不同浓度炭疽菌后发病情况,标尺=10 mm;E-H:利用针刺法对3-4片真叶期辣椒苗接种不同浓度炭疽菌后发病情况,标尺=10 mm;I:果实发病病斑直径统计;J:叶片发病病斑面积统计
Fig. 6 Comparation of fruit needling and leaf needling methods A-D: Pepper fruits inoculated with Colletotrichum spore by using needle point method under different concentration from 1×103 to 1×107 cells/L, bars = 10 mm. E-H: Pepper seedlings inoculated with Colletotrichum spore by using needle point method under different concentration from 1×103 to 1×107 cells/L, bars = 10 mm. I: Diameter of a disease spot on inoculated pepper fruits under each concentration. J: Area of a disease spot on inoculated pepper leaves under each concentration
[1] |
MacNeish RS. Ancient Mesoamerican civilization[J]. Science, 1964, 143(3606): 531-537.
pmid: 17815643 |
[2] | 邹学校, 马艳青, 戴雄泽, 等. 辣椒在中国的传播与产业发展[J]. 园艺学报, 2020, 47(9): 1715-1726. |
Zou XX, Ma YQ, Dai XZ, et al. Spread and industry development of pepper in China[J]. Acta Hortic Sin, 2020, 47(9): 1715-1726.
doi: 10.16420/j.issn.0513-353x.2020-0103 |
|
[3] | 蓝勇. 生活在辣椒时代[J]. 辣椒杂志, 2006, 4(4): 45-49. |
Lan Y. Lived in the Age of chili[J]. J China Capsicum, 2006, 4(4): 45-49. | |
[4] | 周文美. 几种黔式辣椒调味品[J]. 中国调味品, 2001, 26(9): 27-28. |
Zhou WM. Some kinds of chili condiment from Guizhou[J]. Chin Condiment, 2001, 26(9): 27-28. | |
[5] | 李延红, 余顺火. 辣椒碱的提取及其应用研究进展[J]. 农产品加工·学刊, 2006,(10): 71-73. |
Li YH, Yu SH. The progress of research of abstract and application of Capsaicin[J]. The processing of agricultural products, 2006,(10): 71-73. | |
[6] | 王妮, 尹显慧, 彭丽娟, 等. 辣椒炭疽病病原鉴定及其杀菌剂毒力测定[J]. 植物保护, 2019, 45(4): 216-223. |
Wang N, Yin XH, Peng LJ, et al. Identification of the pathogen and toxicity test of fungicides to Capsicum anthracnose[J]. Plant Prot, 2019, 45(4): 216-223. | |
[7] |
毛爱军, 胡洽, 耿三省. 辣椒炭疽病抗病性鉴定技术及利用[J]. 华北农学报, 2004, 19(2): 87-91.
doi: 10.3321/j.issn:1000-7091.2004.02.022 |
Mao AJ, Hu Q, Geng SS. Studies on the inoculation technique for resistant varieties of pepper to anthracnose and the application[J]. Acta Agric Boreali—sinica, 2004, 19(2): 87-91. | |
[8] | 张国芝, 赵霞, 杨海艳, 等. 四川辣椒炭疽病菌鉴定及育种材料抗性筛选[J]. 西南农业学报, 2013, 26(3): 1026-1029. |
Zhang GZ, Zhao X, Yang HY, et al. Identification of pepper anthracnose and resistant screen of breeding materials in Sichuan[J]. Southwest China J Agric Sci, 2013, 26(3): 1026-1029. | |
[9] | 吴庆丽, 秦刚. 辣椒炭疽病抗性资源筛选[J]. 湖北农业科学, 2013, 52(5): 1084-1085, 1089. |
Wu QL, Qin G. Screening on the resistance resources of Capsicum anthracnose[J]. Hubei Agric Sci, 2013, 52(5): 1084-1085, 1089. | |
[10] | 孙春英. 辣椒抗炭疽病QTL定位及分析[D]. 北京: 中国农业科学院, 2013. |
Sun CY. QTL analysis of anthracnose(Colletotrichum acutatum)resistance in Capsicum[D]. Beijing: Chinese Academy of Agricultural Sciences, 2013. | |
[11] | Yoon J, Park HG. Screening method for resistance to pepper fruit anthracnose: pathogen sporulation, inoculation methods related to inoculum concentrations and post-inoculation environment[J]. Horticulture Environment and Biotechnology, 2001, 42(4), PP 389-393 |
[12] | 吴君. 辣椒抗炭疽病材料筛选及抗感材料基因差异表达分析[D]. 武汉: 华中农业大学, 2020. |
Wu J. Screening of anthracnose-resistant pepper germplasms and analysis of differential expression genes in resistant and susceptible materials[D]. Wuhan: Huazhong Agricultural University, 2020. | |
[13] | Abramoff MD, Kardon R, Ts'OD, et al. Intrinsic optical imaging of the retina in awake humans[J]. Invest Ophthalmol Vis. 2004. |
[14] |
Swift M. GraphPad prism, data analysis, and scientific graphing[J]. J Chem Inf Comput Sci, 1997, 37: 411-412.
doi: 10.1021/ci960402j URL |
[15] | 卢鉴植, 刘建华, 巩振辉. 辣椒炭疽病苗期抗病性鉴定方法研究[J]. 江苏农业科学, 1992, 20(5): 42-44. |
Lu JZ, Liu JH, Gong ZH. Research on the method of identifying seedling resistance to pepper anthracnose[J]. Jiangsu Agric Sci, 1992, 20(5): 42-44. | |
[16] | 林清, 吕中华, 黄任中, 等. 辣椒炭疽病抗性鉴定方法研究[J]. 西南农业学报, 2006, 19(6): 1071-1073. |
Lin Q, Lu ZH, Huang RZ, et al. Study on screening method for resistance to pepper anthracnose[J]. Southwest China J Agric Sci, 2006, 19(6): 1071-1073. |
[1] | 赵志祥, 王殿东, 周亚林, 王培, 严婉荣, 严蓓, 罗路云, 张卓. 枯草芽孢杆菌Ya-1对辣椒枯萎病的防治及其对根际真菌群落的影响[J]. 生物技术通报, 2023, 39(9): 213-224. |
[2] | 张蓓, 任福森, 赵洋, 郭志伟, 孙强, 刘贺娟, 甄俊琦, 王童童, 程相杰. 辣椒响应热胁迫机制的研究进展[J]. 生物技术通报, 2023, 39(7): 37-47. |
[3] | 章乐乐, 王冠, 柳凤, 胡汉桥, 任磊. 芒果炭疽病拮抗菌分离、鉴定及生防机制研究[J]. 生物技术通报, 2023, 39(4): 277-287. |
[4] | 徐小文, 李金仓, 海都, 查玉平, 宋菲, 王义勋. 核桃炭疽菌携带病毒种类鉴定及多样性分析[J]. 生物技术通报, 2023, 39(3): 278-289. |
[5] | 杜清洁, 周璐瑶, 杨思震, 张嘉欣, 陈春林, 李娟起, 李猛, 赵士文, 肖怀娟, 王吉庆. 过表达CaCP1提高转基因烟草对盐胁迫的敏感性[J]. 生物技术通报, 2023, 39(2): 172-182. |
[6] | 陈奕博, 杨万明, 岳爱琴, 王利祥, 杜维俊, 王敏. 基于SLAF标记的大豆遗传图谱构建及苗期耐盐性QTL定位[J]. 生物技术通报, 2023, 39(2): 70-79. |
[7] | 段敏杰, 李怡斐, 杨小苗, 王春萍, 黄启中, 黄任中, 张世才. 辣椒锌指蛋白DnaJ-Like基因家族鉴定及对高温胁迫的表达响应[J]. 生物技术通报, 2023, 39(1): 187-198. |
[8] | 石广成, 杨万明, 杜维俊, 王敏. 大豆耐盐种质的筛选及其耐盐生理特性分析[J]. 生物技术通报, 2022, 38(4): 174-183. |
[9] | 胡华冉, 杜磊, 张芮豪, 钟秋月, 刘发万, 桂敏. 辣椒适应非生物胁迫的研究进展[J]. 生物技术通报, 2022, 38(12): 58-72. |
[10] | 唐跃辉, 赵雨凡, 林锦, 王胤, 曹博远, 车怡帆, 杨文杰, 包欣欣, 杨同文. 水稻苗期致死突变体的鉴定及其基因定位[J]. 生物技术通报, 2022, 38(10): 124-131. |
[11] | 刘沙玉, 曹健, 李蒙, 柳志强, 李晓宇. 橡胶树胶孢炭疽菌Zn2Cys6型转录因子CgAswA的生物学功能[J]. 生物技术通报, 2021, 37(9): 161-170. |
[12] | 刘娜, 刘世科, 王倩男. 胶孢炭疽菌细胞骨架荧光标记菌株的构建[J]. 生物技术通报, 2021, 37(8): 284-293. |
[13] | 周静, 黄文茂, 秦利军, 韩丽珍. 四株PGPR菌株混菌发酵体系的构建及促生效应评价[J]. 生物技术通报, 2021, 37(4): 116-126. |
[14] | 杨茉, 高婷, 李滟璟, 魏崇瑶, 高淼, 马莲菊. 辣椒根际促生菌的分离筛选及抗病促生特性研究[J]. 生物技术通报, 2020, 36(5): 104-109. |
[15] | 王姣, 张水, 张婧柔, 邵贵芳, 邓明华. 辣椒胞质雄性不育系CaCOX3的克隆与表达分析[J]. 生物技术通报, 2019, 35(4): 1-6. |
阅读次数 | ||||||
全文 |
|
|||||
摘要 |
|
|||||