新疆棉花茎腐病的病原鉴定及其生物学特性研究

doi:10.11963/1002-7807.sllgy.20210423

摘要/Abstract

摘要：

【目的】自2016年以来在新疆棉田发现1种棉花茎腐病,在潮湿环境下,该病害可导致棉花茎秆上产生纵裂和表皮组织崩溃等腐烂症状;在干燥环境下,病斑扩展明显变慢,甚至不再扩展,呈现坏死斑症状,严重时会导致茎秆枯死。明确其病原种类及生物学特性对病害防治至关重要。【方法】从南北疆11个地点采集病样18份。采用组织分离法分离病原,经单孢纯化,共获得20株代表性菌株。利用形态学、分子生物学方法对病原进行种的鉴定,并测定菌株的致病性及其代表性菌株在不同温度、光照、pH和培养基条件下的生物学特性和致死温度,了解适宜生长、产孢条件。【结果】引起新疆棉花茎腐病病原为Fusarium incarnatum和F. proliferatum,其中F. incarnatum为优势种。适于2种病原菌生长和产孢的温度均为25 ℃;光照有利于F. incarnatum生长,黑暗培养有利于二者产孢。二者生长的最适pH均为8,产孢的最适pH均为7。在马铃薯蔗糖琼脂培养基、马铃薯葡萄糖琼脂培养基上,二者生长和产孢较好。F. incarnatum和F. proliferatum孢子的致死温度分别为48 ℃、60 ℃,菌丝的致死温度分别为50 ℃、61 ℃。【结论】引起新疆棉花茎腐病的病原菌为F. incarnatum和F. proliferatum,其中F. incarnatum为优势种。20~30 ℃、pH 7~8和全光照条件有利于其生长,黑暗条件有利于其产孢,马铃薯蔗糖琼脂培养基和马铃薯葡萄糖培养基均有利于其生长和产孢。

关键词: 棉花; 茎腐病; 病原鉴定; 生物学特性; 培养条件; 致死温度

Abstract:

[Objective] A cotton stem rot disease has been observed in Xinjiang Uygur Autonomous Region of China since 2016. In wet conditions, part of the diseased stem can produce rot symptoms such as longitudinal cracks and collapse of epidermal tissue; in dry conditions, the expansion of the diseased spots is significantly slower, or even no longer expands, showing symptoms of necrotic spots. In severe cases, the stems will die. Hence, it is vital to understand the species and biological characteristics of pathogens for disease control. [Method] A total of 18 stems of cotton with symptoms rot were collected from 11 cotton planting locations in the north and south of Xinjiang Uygur Autonomous Region. The pathogens were isolated and purified by conventional dilution separation and 20 representative strains were obtained by single spore purification. The pathogens were identified by morphology, molecular biology, and pathogenicity. To understand the suitable conditions for the growth and sporulation of the pathogens, the biological characteristics of the pathogens such as different temperatures, light, pH values, media and lethal temperatures were tested. [Result]The pathogens of cotton stem rot in Xinjiang were Fusarium incarnatum and F. proliferatum, among which F. incarnatum was the dominant species. The suitable temperature for the growth and sporulation of the two pathogens are 25 ℃. Light is beneficial to the growth of F. incarnatum, and dark culture is beneficial to the sporulation of two pathogens. The optimal pH for the growth of these two pathogens is 8, and the optimal pH for sporulation is 7. These two kinds of pathogens grow and produce spores better on potato sucrose agar and potato dextrose agar. The lethal temperatures of F. incarnatum and F. proliferatum spores were 48 ℃ and 60 ℃, respectively, and the lethal temperatures of hyphae were 50 ℃ and 61 ℃, respectively. [Conclusion] The pathogens of cotton stem rot in Xinjiang were F. incarnatum and F. proliferatum. F. incarnatum was identified as the dominant species. It is beneficial to the growth of the two species of Fusarium under the conditions of 20-30 ℃, pH 7-8 and full light; and the spore will be more productive under dark. Both potato sucrose agar and potato dextrose agar medium are beneficial to the growth and sporulation of the two pathogens.

Key words: cotton; stem rot; pathogen identification; biological characteristics; cultural condition; lethal temperature

孙璘,海艳,唐晓雪,祖丽皮亚·艾买,焦瑞莲,任毓忠,李国英. 新疆棉花茎腐病的病原鉴定及其生物学特性研究[J]. 棉花学报, 2021, 33(3): 235-246.

Sun Lin,Hai Yan,Tang Xiaoxue,Zulipiya Aimai,Jiao Ruilian,Ren Yuzhong,Li Guoying. Identification and biological characteristics of the pathogens of cotton stem rot in Xinjiang[J]. Cotton Science, 2021, 33(3): 235-246.

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链接本文: http://journal.cricaas.com.cn/Jweb_mhxb/CN/10.11963/1002-7807.sllgy.20210423

http://journal.cricaas.com.cn/Jweb_mhxb/CN/Y2021/V33/I3/235

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样品编号 Sample number	采样地点 Collection location	采集时间（年-月） Collecting time (year-month)
F1	阿拉尔地区新疆生产建设兵团（简称“新疆兵团”）第一师10团10th Regimental Farm of First Division, Xinjiang Production and Construction Corps (Xinjiang Crops)	2017-08
F2	阿拉尔塔河种业实验场 Experimental Field of Tahe Seed Industry in Aral	2017-08
F3	阿拉尔地区新疆兵团第一师12团 12th Regimental Farm of First Division of Xinjiang Corps in Aral	2017-08
F4~F5	石河子大学实验场Teaching Experiment Field of Shihezi University	2017-08
F6~F7	石河子地区新疆兵团第八师150团 150th Regimental Farm, Eighth Division, Xinjiang Corps	2017-08
F8	新疆兵团第八师石河子总场4分场3连 Company 3, 4th Branch of the General Farm of Eighth Division of Xinjiang Crops in Shihezi	2017-08
F9	石河子地区新疆兵团第八师133团 133th Regimental Farm, Eighth Division of Xinjiang Corps in Shihezi	2017-08
F10~F13	新疆兵团第八师石河子总场3分场3连 Company 3, 3rd Branch of the General Farm of Eighth Division of Xinjiang Corps in Shihezi	2020-09
F14	石河子地区新疆兵团第八师143团3分场3连 Company 3, 3rd Branch of the 143 Regimental Farm, Eighth Division of Xinjiang Corps in Shihezi	2020-09
F15	新疆农垦科学院 Xinjiang Uygur Autonomous Region Academy of Agricultural Sciences	2020-09
F16~F20	博尔塔拉蒙古自治州精河县大河沿子镇 Daheyanzi Town of Jinghe County in Bortala Mongolian Autonomous Prefecture	2020-09

目标区域 Target region	引物 Primer	引物序列（5'-3'） Primer sequence (5'-3')	退火温度Annealing temperature/℃	参考文献 References
Fusarium	ITSF	AACTCCCAAACCCCTGTGAACATA	58	[11]
spp. ITS	ITSR	TTTAACGGCGTGGCCGC
rRNA-ITS	ITS1	TCCGTAGGTGAACCTGCGG	58	[12]
	ITS4	TCCTCCGCTTATTGATATGC
TEF-1α	TEF-lαF	ATGGGTAAGGAGGACAAGAC	58	[13]
	TEF-1αR	GGAAGTACCAGTGATCATGTT

菌株号 Strain number	片段长度 Fragment length/bp	登录号 Accession number	菌株号 Strain number	片段长度 Fragment length/bp	登录号 Accession number
F1	532	MW659168.1	F15	520	MW659182.1
F2	531	MW659169.1	F16	520	MW659183.1
F3	532	MW659170.1	F17	521	MW659184.1
F4	532	MW659171.1	F18	520	MW659185.1
F5	521	MW659172.1	F19	520	MW659186.1
F6	521	MW659173.1	F20	520	MW659187.1
F7	532	MW659174.1	CBS132194	524	KF255427.1
F8	532	MW659175.1	CBS132317	524	KF255432.1
F9	531	MW659176.1	NRRL13379	1132	GQ505680.1
F10	532	MW659177.1	NRRL32866	1131	GQ505704.1
F11	520	MW659178.1	CBS131574	483	JX162373.1
F12	520	MW659179.1	CBS480.77	407	KR071678.1
F13	522	MW659180.1	CBS125014	407	KR071679.1
F14	520	MW659181.1	CBS131785	483	JX162388.1

菌株号 Strain number	片段长度 Fragment length/bp	登录号 Accession number	菌株号 Strain number	片段长度 Fragment length/bp	登录号 Accession number
F1	684	MW695504.1	F15	678	MW695518.1
F2	684	MW695505.1	F16	676	MW695519.1
F3	684	MW695506.1	F17	676	MW695520.1
F4	684	MW695507.1	F18	676	MW695521.1
F5	676	MW695508.1	F19	676	MW695522.1
F6	676	MW695509.1	F20	678	MW695523.1
F7	684	MW695510.1	CBS132194	680	KF255470.1
F8	684	MW695511.1	CBS132317	680	KF255476.1
F9	684	MW695512.1	NRRL13379	681	GQ505591.1
F10	684	MW695513.1	NRRL32866	681	GQ505615.1
F11	676	MW695514.1	CBS131574	699	JX118983.1
F12	676	MW695515.1	CBS480.77	519	KR071736.1
F13	676	MW695516.1	CBS125014	519	KR071738.1
F14	676	MW695517.1	CBS131785	698	JX118998.1

因素 Factors	试验参数 Parameter	F. incarnatum		F. proliferatum
因素 Factors	试验参数 Parameter	菌落直径 Colony diameter / mm	产孢量 Sporulation/ （10⁶ mL^-1）	菌落直径 Colony diameter / mm	产孢量 Sporulation/ （10⁶ mL^-1）
温度	5	0.00±0.00 f	0.00±0.00 d	0.00±0.00 f	0.00±0.00 e
Temperature/℃	10	9.68±0.79 e	0.00±0.00 d	10.45±0.43 d	0.15±0.05 de
	15	18.82±0.02 d	0.08±0.02 c	19.87±0.58 c	0.51±0.03 c
	20	50.93±0.39 b	0.28±0.01 b	48.32±0.93 b	0.59±0.06 c
	25	62.62±0.88 a	1.40±0.01 a	60.28±1.07 a	2.56±0.20 a
	30	45.23±1.79 c	1.39±0.01 a	48.32±2.15 b	1.16±0.03 b
	35	0.00±0.00 f	0.00±0.00 d	7.45±0.25 e	0.24±0.06 d
光照	完全光照Light (L)	70.55±1.01 a	0.01±0.00 c	60.23±0.73 a	1.72±0.14 a
Light condition	完全黑暗Dark (D)	65.28±2.16 b	1.14±0.03 a	60.08±0.47 a	2.45±0.72 a
	光暗交替Alternating L & D	68.18±2.29 ab	0.23±0.03 b	59.90±0.01 a	1.66±0.11 a
pH	4	34.98±1.74d	0.68±0.07 d	45.92±0.98 c	0.80±0.04 cd
	5	54.40±0.94 c	1.12±0.06 ab	55.57±1.79 b	0.94±0.09 c
	6	59.70±0.92 b	1.15±0.33 ab	56.63±0.94 b	0.94±0.03 c
	7	60.33±0.85 b	1.33±0.17 a	62.98±0.51 a	1.80±0.12 a
	8	63.27±1.69 a	1.04±0.00 bc	64.97±0.99 a	1.31±0.11 b
	9	62.28±0.44 ab	0.96±0.05 bc	63.98±0.52 a	0.72±0.09 d
	10	60.52±1.90 ab	0.82±0.01 cd	63.97±0.12 a	0.70±0.04 d
培养基	PSA	65.92±0.88 ab	1.27±0.07 a	64.35±7.26 a	2.19±0.29 b
Culture media	PDA	67.48±0.47 a	1.17±0.03 a	61.88±1.67 a	2.69±0.13 a
	CMA	48.86±1.23 d	0.01±0.00 d	52.21±2.74 cd	0.03±0.00 d
	MEA	64.41±1.00 b	0.42±0.01 c	50.52±0.68 cd	2.07±0.43 b
	OA	59.07±0.67 c	0.45±0.18 c	53.93±0.35 bc	0.72±0.03 c
	VBC	40.90±2.88 e	1.08±0.10 ab	48.77±2.18 d	0.93±0.09 c
	SNA	43.38±0.68 e	0.83±0.03 b	56.08±1.98 b	0.64±0.08 c