辣椒苗期炭疽菌接种方法

doi:10.13560/j.cnki.biotech.bull.1985.2022-0994

摘要/Abstract

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。

关键词: 辣椒, 苗期, 炭疽菌, 接种方法

Abstract:

Anthracnose is one of the most serious diseases in pepper production, there is an urgent need for reliable methods to screen for Colletotrichum disease resistance in the early pepper development. In this study, by using Colletotrichum strain CD, we have introduced two methods to inoculate pepper ‘ST-8’ seedling: infiltration and needle point, and tried to find the optimal way by comparing the plant infection rate, leave curl rate and the size of necrotic disease spots. The results showed that the results obtained by using the needle point method under the resuspend spore concentration of 1 × 10⁶cells/mL were the best, which is expected to be applied to large-scale screening of pepper being resistant to anthracnose.

Key words: Capsicum annuum L., seedling, Colletotrichum, inoculation method

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/表 7

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

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 1^st - 2^nd leave		3-4片真叶期 3^rd - 4^th leave				5-10片真叶期 5^th - 10^thleave		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

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