云南省绿春县胡椒炭疽病菌的分离与鉴定

doi:10.16178/j.issn.0528-9017.20240360

摘要/Abstract

摘要：

炭疽病是胡椒叶片上的一种常见病害，严重时会导致植株叶片枯萎脱落，影响胡椒产量，造成一定的经济损失。炭疽菌属（Colletotrichum）真菌是引起胡椒（Piper nigrum L.）枝叶枯死的重要病原菌。绿春县是云南省的胡椒主产区，当地长年的湿、热气候导致胡椒炭疽病发生频繁。本研究从患病组织中分离、纯化得到病原菌H4，并对其进行致病性测定。根据形态特征和内转录间隔区（internal transcribed spacer，ITS）序列、肌动蛋白基因（Actin）、甘油醛-3-磷酸脱氢酶基因（GADPH）联合构建系统发育树分析，明确引起胡椒叶片炭疽病的病原菌为暹罗炭疽菌（Colletotrichum siamense），这是首次在云南省内发现的由暹罗炭疽菌引起的胡椒叶片炭疽病，研究结果为绿春县胡椒病害的绿色防控技术提供了理论依据。

关键词: 炭疽病, 胡椒, 病原菌, 鉴定, 暹罗炭疽菌

Abstract:

Anthracnose is a common disease on pepper leaves，which can cause plant leaves to wither and fall-off in severe cases，affecting pepper yield and causing certain economic losses. Colletotrichum fungi are important pathogens that cause the withering of pepper branches and leaves. Lyuchun County is the main pepper production area in Yunnan Province，and due to the local humid and hot climate，pepper anthracnose occurs frequently. This study isolated and purified a pathogenic fungus strain H4 from diseased tissues，and determined its pathogenicity. Based on morphological characteristics and the combined construction of internal transcribed spacer （ITS） sequence， Actin gene，and GADPH gene，a phylogenetic tree analysis was conducted，and it was determined that the pathogen causing anthracnose in pepper leaves was Colletotrichum siamense. To our knowledge，this was the first discovery of anthracnose on pepper leaves in Yunnan caused by C. siamense. And the relevant results provide important theoretical basis for the prevention and control of pepper diseases.

Key words: anthracnose, pepper（Piper nigrum L.）, pathogen, identification, Colletotrichum siamense

中图分类号:

S435.621.2⁺2

和国优, 张义杰, 时春月, 鲁耀, 伏成秀, 王振充, 严钦, 孔垂思. 云南省绿春县胡椒炭疽病菌的分离与鉴定[J]. 浙江农业科学, 2026, 67(2): 444-448.

HE Guoyou, ZHANG Yijie, SHI Chunyue, LU Yao, FU Chengxiu, WANG Zhenchong, YAN Qin, KONG Chuisi. Isolation and identification of pepper anthracnose pathogen in Lyuchun County，Yunnan Province of China[J]. Journal of Zhejiang Agricultural Sciences, 2026, 67(2): 444-448.

图/表 5

表 1 病原菌鉴定所用引物

Table 1 Primers used for pathogen identification

基因	引物	序列（5′→3′）	参考文献
ITS	ITS1	TCCGTAGGTGAACCTGCGG	［9］
ITS	ITS2	TCCTCCGCTTATTGATATGC	［9］
Actin	ACT-512F	ATGTGCAAGGCCGGTTTCGC	［9］
Actin	ACT-783R	TACGAGTCCTTCTGGCCCAT	［9］
GAPDH	GDF1	GCCGTCAACGACCCCTTCATTGA	［9］
GAPDH	GDR1	GGGTGGAGTCGTACTTGAGCATGT	［9］

图1 胡椒叶炭疽病的症状A—自然感染叶片；B—人工接种叶片。

Fig.1 Symptoms of Piper nigrum L. leaf anthracnose

图2 PDA平板上的菌落A—菌落（6 d，正面）；B—反向菌落（6 d，反面）；C—菌落（30 d）；D—分生孢子堆（20 d）。

Fig.2 Morphological appearances of colonies on PDA plates

图3 炭疽病病原菌 C. siamense 的形态学特征A—菌丝体（20×）；B—菌丝体（40×）；C—分生孢子（40×）；D—分生孢子（100×）。

Fig.3 Morphological characteristics of the pathogen of C. siamense

图4 基于ITS、Actin和GAPDH基因的Colletotrichum系统发育树

Fig.4 Phylogenetic tree of Colletotrichum based on combined ITS，Actin and GAPDH genes

参考文献 20

[1]	RAVINDRAN P N. Black pepper：Piper nigrum ［M］. CRC Press，2000.
[2]	胡丽松. “香料之王” 胡椒［J］. 生命世界，2020（8）：24-25.
	HU L S. “King of spices” pepper［J］. Life World，2020（8）：24-25.（in Chinese）
[3]	HU L S， XU Z P， WANG M J，et al. The chromosome-scale reference genome of black pepper provides insight into piperine biosynthesis［J］. Nature Communications，2019，10（1）：4702.
[4]	LIU S C， LIU R B， CHU B，et al. Identification and screening of fungicides against Piper nigrum basal Fusarium wilt disease in Hainan，China［J］. Journal of Basic Microbiology，2023，63（11）：1254-1264.
[5]	沈绍斌. 胡椒：云南省特色产业［J］. 中国农村科技，2020（9）：74-75.
	SHEN S B. Pepper：a characteristic industry in Yunnan Province［J］. China Rural Science & Technology，2020（9）：74-75.
[6]	段春芳，黄贵修，郭容琦，等. 云南胡椒炭疽病病原鉴定［J］. 江西农业学报，2012，24（11）：56-58.
	DUAN C F， HUANG G X， GUO R Q，et al. Identification of pathogen of anthracnose in pepper（Pipper nigrum）［J］. Acta Agriculturae Jiangxi，2012，24（11）：56-58.
[7]	CANNON P F， DAMM U， JOHNSTON P R，et al. Colletotrichum：current status and future directions［J］. Studies in Mycology，2012，73：181-213.
[8]	LIN C H， FU L X， FAN Z Z，et al. First report of leaf anthracnose caused by Colletotrichum karstii of Piper nigrum in Yunnan Province，China［J］. Plant Disease，2022，106（4）：1303.
[9]	VERMA R，DAS A， CHAKRAWARTI N，et al. First report of black pepper （Piper nigrum） anthracnose caused by Colletotrichum siamense in north-east India［J］. Plant Disease，2023，107（7）：2249.
[10]	DAMM U， CANNON P F， WOUDENBERG J H C，et al. The Colletotrichum acutatum species complex［J］. Studies in Mycology，2012，73：37-113.
[11]	MARIN-FELIX Y， GROENEWALD J Z， CAI L，et al. Genera of phytopathogenic fungi：GOPHY 1［J］. Studies in Mycology，2017，86：99-216.
[12]	WALKER J. The coelomycetes：fungi imperfecto with pycnidia，acervuli and stromata［J］. Australasian Plant Pathology，1980（9）：120-121.
[13]	DE SILVA D D， GROENEWALD J Z， CROUS P W，et al. Identification，prevalence and pathogenicity of Colletotrichum species causing anthracnose of Capsicum annuum in Asia［J］. IMA Fungus，2019，10：8.
[14]	JAYASINGHE C K， FERNANDO T H P S， JAYAWARDANA N. A comparative study of Colletotrichum species causing anthracnose in Hevea［J］. Journal of the Rubber Research Institute of Sri Lanka，2009，89：20-32.
[15]	XU M， ZHOU Y， ZHANG R，et al. First report of leaf spot caused by Colletotrichum siamense on Nageia nagi in China［J］. Plant Disease，2023，107（3）：961.
[16]	PÉREZ-MORA J L， MORA-ROMERO G A， BELTRÁN-PEÑA H，et al. First report of Colletotrichum siamense and C. gloeosporioides causing anthracnose of Citrus spp. in Mexico［J］. Plant Disease，2021，105（2）：496.
[17]	WANG Q H， XU M， QI Y K，et al. Morphological and molecular identification of Colletotrichum siamense associated with Cayratia japonica anthracnose in China［J］. Crop Protection，2023，174：106385.
[18]	HERATH I S， UDAYANGA D， MIRIYAGALLA S，et al. Colletotrichum siamense causing anthracnose-twister disease of onion （Allium cepa） in Sri Lanka［J］. Australasian Plant Disease Notes，2021，16：30.
[19]	ZAKARIA L. Diversity of Colletotrichum species associated with anthracnose disease in tropical fruit crops：a review［J］. Agriculture，2021，11（4）：297.
[20]	SANDERS G， KORSTEN L. A comparative morphology of South African avocado and mango isolates of Colletotrichum gloeosporioides ［J］. Canadian Journal of Botany，2011，81，877-885.