Inoculation Method for Colletotrichum in Pepper（Capsicum annuum）Seedlings

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

Abstract

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

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.

Figures/Tables 7

Fig. 1 Needle point method A: During inoculation（arrowheads: needle point position）. B: After inoculation（arrowheads: inoculated position）

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

Fig. 3 Batch measurement of lesion area by using ImageJ A: Set up photo scale. B: Choose lesion area. C: Measure chosen areas

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

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

Fig. 5 Colletotrichum inoculated leaves by using needle point method during different developmental stages of pepper seedlings at different Colletotrichum spore concentration

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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