Investigation and Species Identification of Cyst Nematode Disease on Rice in Guangdong Province

doi:10.16819/j.1001-7216.2024.231001

Abstract

Abstract:

【Objective】 To investigate the species, distribution, and occurrence of cyst nematode disease on rice in Guangdong Province. 【Methods】 A total of 231 rice roots and rhizosphere soil samples from 14 cities in Guangdong Province were collected and analyzed. The cyst nematode species in these samples were identified through morphological and molecular identification. 【Results】 Cyst nematodes were found on rice in Qingyuan, Zhaoqing, and Jiangmen in Guangdong Province, with detection rates of 40%, 23.7%, and 10%, respectively. Morphological identification revealed that the cysts were lemon to spherical-shaped, predominantly lemon-shaped, with an ambifenestrate vulval cone, a nearly round fenestral, a narrow vulval bridge, and a medium-sized underbridge. The second-stage juvenile (J2) had a hemispherical and offset labial region, a conical tail with a pointed tip, and a hyaline region occupying 53% to 66% of the tail length. Molecular identification indicated that the 28S rDNA D2-D3 region of the cyst nematode was 99.73%－100% similar to Heterodera elachista in the NCBI database, and the rDNA-ITS region was 96.72%－99.91% similar to Heterodera elachista in the NCBI database. Phylogenetic analysis placed the nematode in the same branch as H. elachista with a confidence level of 100%. Using specific primers He-F/He-R of H. elachista, a 281 bp single band was amplified from the rDNA-ITS region of the cyst nematodes, consistent with previous reports. 【Conclusion】 The cyst nematodes collected in Qingyuan, Zhaoqing, and Jiangmen were identified as H. elachista, marking the first report of H. elachista in Guangdong Province.

Key words: Heterodera elachista, rice, occurrence and distribution, species identification

摘要：

【目的】 明确广东省水稻上孢囊线虫发生种类、分布。【方法】 从广东省的14个市采集了231份水稻根系和根际土壤样品，从水稻根系及根际土壤样品中分离孢囊线虫，采用形态学和分子生物学方法鉴定孢囊线虫种类。【结果】 在广东清远、肇庆、江门等地水稻根系及根际土样中发现孢囊线虫，孢囊检出率分别为40%、23.7%、10%。形态学研究表明，孢囊柠檬形或近圆形，主要为柠檬形，阴门锥双半膜孔，膜孔近圆形，阴门桥窄，下桥中等大小；二龄幼虫唇区半球形，略缢缩；尾圆锥形，尾端尖，透明尾长占尾长的 53%~66%。分子生物学研究表明，该线虫28S rDNA D2-D3区序列与NCBI数据库中旱稻孢囊线虫相似度99.73%~100%，rDNA-ITS区序列与NCBI数据库中旱稻孢囊线虫相似度96.72%~99.91%。系统进化树分析表明，该线虫与旱稻孢囊线虫处于同一分支，置信度为100%。使用旱稻孢囊线虫特异性引物He-F/He-R扩增清远、肇庆、江门采集的孢囊线虫rDNA-ITS区均得到281 bp 的单一条带，与已报道结果一致。【结论】 清远、肇庆、江门采集到的孢囊线虫均为旱稻孢囊线虫。这是首次在广东省发现旱稻孢囊线虫。

关键词: 旱稻孢囊线虫, 水稻, 发生分布, 种类鉴定

LIU Fuxiang, ZHEN Haoyang, PENG Huan, ZHENG Liuchun, PENG Deliang, WEN Yanhua. Investigation and Species Identification of Cyst Nematode Disease on Rice in Guangdong Province[J]. Chinese Journal OF Rice Science, 2024, 38(4): 456-461.

刘福祥, 甄浩洋, 彭焕, 郑刘春, 彭德良, 文艳华. 广东省水稻孢囊线虫病调查与鉴定[J]. 中国水稻科学, 2024, 38(4): 456-461.

Figures/Tables 6

References 28

[1]	朱德峰, 张玉屏, 陈惠哲, 向镜, 张义凯. 中国水稻高产栽培技术创新与实践[J]. 中国农业科学, 2015, 48(17): 3404-3414.
	Zhu D F, Zhang Y P, Chen H Z, Xiang J, Zhang Y K. Innovation and practice of high-yield rice cultivation technology in China[J]. Scientia Agricultura Sinica, 2015, 48(17): 3404-3414. (in Chinese with English abstract)
[2]	Pokharel R R, Abawi G S, Zhang N, Duxbury J M, Smarti C D. Characterization of isolates of Meloidogyne from rice-wheat production fields in nepal[J]. Nematolog, 2007, 39(3): 221-223.
[3]	谢家廉, 杨芳, 黄文坤, 彭德良, 彭云良, 姬红丽. 近年水稻主要线虫病害的研究进展[J]. 植物保护学报, 2017, 44(6): 940-949.
	Xie J L, Yang F, Huang W K, Peng D L, P Y L, Ji H L. Advances in major rice parasitic nematodes in recent years[J]. Journal of Plant Protection, 2017, 44(6): 940-949. (in Chinese with English abstract)
[4]	丁中, Namphueng J, 何旭峰, 伍敏敏, 洪宏. 旱稻孢囊线虫生活史及侵染特性[J]. 中国水稻科学, 2012, 26(6): 746-750.
	Ding Z, Namphueng J, He X F, Wu M M, Hong H. Life cycle and infection characteristics of rice cyst nematode, Heterodera elachista Ohshima in rice[J]. Chinese Journal of Rice Science, 2012, 26(6): 746-750. (in Chinese with English abstract)
[5]	卓侃, 宋汉达, 王宏洪, 陶冶, 张洪玲, 陆秀红, 黄金玲, 刘志明, 廖金铃. 旱稻孢囊线虫在广西的发生及其rDNA-ITS异质性分析[J]. 中国水稻科学, 2014, 28(1): 78-84.
	Zhuo K, Song H D, Wang H H, Tao Y, Zhang H L, Lu X H, Huang J L, Liu Z M, Liao J L. Occurrence of Heterodera elachista in Guangxi region and its intra-species heterogeneity in rDNA-ITS region[J]. Chinese Journal of Rice Science, 2014, 28(1): 78-84. (in Chinese with English abstract)
[6]	Ding Z, Namphueng J, He X F, Peng D L, Huang W K. First report of the cyst nematode (Heterodera elachista) on rice in Hunan Province, China[J]. Plant Disease, 2012, 96(1): 151.
[7]	Cui J K, Zhou B, Jiao Y J, Chen K Y, Ren H H, Meng H G, Jiang S J. First report of Heterodera elachista infecting rice in Henan Province of Central China[J]. Plant Disease, 2021, 105(2): 511.
[8]	崔思佳, 张大帆, 彭德良, 黄文坤, 丁中. 旱稻孢囊线虫的发生分布及寄生调查[C]. 中国植物保护学会.病虫害绿色防控与农产品质量安全——中国植物保护学会2015年学术年会论文集. 北京: 中国农业科学技术出版社, 2015.
	Cui S J, Zhang D F, Peng D L, Huang W K, Ding Z. Investigation on occurrence, distribution and parasitism of Heterodera elachista[C]. Green Pest and Disease Control and Agricultural Product Quality Safety- Proceedings of Chinese Society of Plant Protection, Beijing: China Agricultural Science and Technology Press, 2015. (in Chinese with English abstract)
[9]	甄浩洋. 赣粤两省作物孢囊线虫调查及茶皂素颗粒剂对其防治研究[D]. 广州: 华南农业大学, 2018.
	Zhen H Y. Survey of cyst-forming nematodes in Jiangxi and Guangdong provinces and field control of cyst-forming nematodes using tea saponin granules[D]. Guangzhou: South China Agricultural University, 2018. (in Chinese with English abstract)
[10]	Xiao D X, Feng Y X, Zhou Y, Xuan Y H, Chen L J, Wang Y Y, Liu X Y, Duan Y X, Zhu X F. First report of Heterodera elachista on Zea mays and Echinochloa crusgalli var. mitis in Liaoning Province, China[J]. Plant Disease, 2019, 103(6): 1433.
[11]	Xu Z P, Li H X, Liu Y G, Li W H, Zhang S L. First report of cyst nematode on Zea mays in Gansu Province, China[J]. Plant Disease, 2021, 105(2): 511.
[12]	任豪豪, 陈昆圆, 周博, 焦永吉, 蒋士君, 汤继华, 崔江宽. 河南省禾谷类作物孢囊线虫的发生分布与种类鉴定[J]. 植物保护学报, 2022, 49(6): 1685-1696.
	Ren H H, Chen K Y, Zhou B, Jiao Y J, Jiang S J, Tang J H, Cui J K. Occurrence and identification of cereal crops cyst nematode in Henan Province of China[J]. Journal of Plant Protection, 2022, 49(6): 1685-1696. (in Chinese with English abstract)
[13]	Bridge J, Luc M, Plowright R A. Nematode parasites of rice[M]. Wallingford, UK: CAB International, 1990.
[14]	黄勇椿, 叶姗, 袁涛, 周建宇, 彭德良, 黄文坤, 丁中. 湖南长沙一季晚稻旱稻孢囊线虫发生动态及危害损失[J]. 中国水稻科学, 2021, 35(1): 98-102.
	Huang Y C, Ye S, Yuan T, Zhou J Y, Peng D L, Huang W K, Ding Z. Population dynamics and yield loss evaluation of Heterodera elachista in single-cropping late rice in Changsha, China[J]. Chinese Journal of Rice Science, 2021, 35(1): 98-102. (in Chinese with English abstract)
[15]	Shimizu K. Influence of the upland rice cyst nematode, Heterodera elachista, on the yield of the upland paddy rice[J]. Japanese Journal of Nematology, 1976, 6: 1-6.
[16]	Ito T, Araki M, Komatsuzaki M. No-tillage cultivation reduces rice cyst nematode (Heterodera elachista) in continuous upland rice (Oryza sativa) culture and after conversion to soybean (Glycine max) in Kanto, Japan[J]. Field Crops Research, 2015, 179: 44-51.
[17]	潘阳阳, 黄道强, 王重荣, 王志东, 李宏, 周德贵, 陈宜波, 赵雷, 龚蓉周少川. 广东省近40年常规水稻审定品种特性分析[J]. 中国稻米, 2023, 29(3): 74-79.
	Pan Y Y, Huang D Q, Wang C R, Wang Z D, Li H, Zhou D G, Chen Y B, Zhao L, Gong R, Zhou S C. Analysis on the characteristics of approved conventional rice varieties in Guangdong province in the past 40 years[J]. China Rice, 2023, 29(3): 74-79. (in Chinese with English abstract)
[18]	李怡珍, 金殷文, 陈纯. 水稻孢囊线虫鉴定初报[J]. 植物检疫, 1985, (1): 54-57.
	Li Y Z, Jin Y W, Chen C. Preliminary report of Heterodera oryzicola identification[J]. Plant Quarantine, 1985, (1): 54-57. (in Chinese)
[19]	冯志新. 植物线虫学[M]. 北京: 中国农业出版社, 2001.
	Feng Z X. Plant Nematology[M]. Beijing: China Agriculture Press, 2001. (in Chinese)
[20]	邓春菊, 李艳, 杨毅娟, 杨艳梅, 陈敏, 李永青, 胡先奇. 云南省马铃薯孢囊线虫种类鉴定[J]. 云南农业大学学报, 2023, 38(3): 400-408.
	Deng C J, Li Y, Yang Y J, Yang Y M, Chen M, Li Y Q, Hu X Q. Identification of potato cyst nematodes (Globodera rostochiensis) on potato in Yunnan Province[J]. Journal of Yunnan Agricultural University, 2023, 38(3): 400-408. (in Chinese with English abstract)
[21]	谢辉. 植物线虫分类学[M]. 北京: 高等教育出版社, 2005.
	Xie H. Taxonomy of plant nematodes[M]. Beijing: Higher Education Press, 2005. (in Chinese)
[22]	Subbotin S A, Mundo-Ocampo M, Baldwin J G, Systematics of cyst nematodes (Nematoda:Heteroderinae)//Hunt D J, Perry R N. ed. Nematology Monographs and Perspectives 8A[M]. Leiden, The Netherlands: BRILL, 2010.
[23]	王水南, 彭德良, 黄文坤, 丁中. 旱稻孢囊线虫的快速分子检测[J]. 湖南农业大学学报, 2014, 40(2): 178-182.
	Wang S N, Peng D L, Huang W K, Ding Z. Rapid molecular diagnosis of rice cyst nematode (Heterodera elachista)[J]. Journal of Hunan Agricultural University, 2014, 40(2): 178-182. (in Chinese with English abstract)
[24]	Minh B Q, Hahn M W, Lanfear R. New methods to calculate concordance factors for phylogenomic datasets[J]. Molecular Biology and Evolution, 2020, 37, 2727-2733.
[25]	Luca F D, Vovlas N, Lucarelli G, Troccoli A, Radicci V, Fanelli E, Cantalapiedra-Navarrete C, Palomares-Rius J E, Castillo P. Heterodera elachista the Japanese cyst nematode parasitizing corn in Northern Italy: Integrative diagnosis and bionomics[J]. European Journal of Plant Pathology, 2013, 136(4): 857-872.
[26]	Ohshima Y. Heterodera elachista n. sp., an upland rice cyst nematode from Japan[J]. Japanese Journal of Nematology, 1974, 4: 51-56.
[27]	Nobbs J, Ibrahim S K, Rowe J. A morphological and biochemical comparison of the four cyst nematode species, Heterodera elachista, H. oryzicola, H. oryzae and H. sacchari (Nematoda: Heteroderidae) known to attack rice (Oryza sativa). Fundamental and Applied Nematology, 1992, 15, 551-562.
[28]	Tanha M Z, Subbotin S A, Moens M. Molecular identification of cyst-forming nematodes (Heteroderidae) from Iran and a phylogeny based on ITS-rDNA sequences[J]. Nematology, 2003, 5, 99-111.

调查地点 Site of investigation	寄主 Host	采集样品数 Number of samples	孢囊样品数 Number of cyst samples	孢囊检出率 Detection rate of cysts(%)
清远Qingyuan	水稻Rice	25	10	40
肇庆Zhaoqing	水稻Rice	38	9	23.7
江门Jiangmen	水稻Rice	30	3	10
惠州Huizhou	水稻Rice	1	0	0
揭阳Jieyang	水稻Rice	2	0	0
广州Guangzhou	水稻Rice	3	0	0
梅州Meizhou	水稻Rice	4	0	0
阳江Yangjiang	水稻Rice	4	0	0
河源Heyuan	水稻Rice	5	0	0
汕尾Shanwei	水稻Rice	5	0	0
韶关Shaoguan	水稻Rice	21	0	0
云浮Yunfu	水稻Rice	24	0	0
茂名Maoming	水稻Rice	26	0	0
湛江Zhanjiang	水稻Rice	43	0	0

调查地点 Site of investigation	寄主 Host	采集样品数 Number of samples	孢囊样品数 Number of cyst samples	孢囊检出率 Detection rate of cysts(%)
清远Qingyuan	水稻Rice	25	10	40
肇庆Zhaoqing	水稻Rice	38	9	23.7
江门Jiangmen	水稻Rice	30	3	10
惠州Huizhou	水稻Rice	1	0	0
揭阳Jieyang	水稻Rice	2	0	0
广州Guangzhou	水稻Rice	3	0	0
梅州Meizhou	水稻Rice	4	0	0
阳江Yangjiang	水稻Rice	4	0	0
河源Heyuan	水稻Rice	5	0	0
汕尾Shanwei	水稻Rice	5	0	0
韶关Shaoguan	水稻Rice	21	0	0
云浮Yunfu	水稻Rice	24	0	0
茂名Maoming	水稻Rice	26	0	0
湛江Zhanjiang	水稻Rice	43	0	0

形态特征 Morphological character	广东清远 Qingyuan, Guangdong, Province, China(This study)	日本 Japan(1974)^[26]	日本 Japan (1992) ^[27]	湖南 Hunan, China (2012) ^[6]	广西 Guangxi, China (2014)^[5]
孢囊(测量标本数)cysts(n)	20	10	50	20	5
体长(不含颈)Body length(excl. neck) (L)	484.90±41.80 (386.00~542.00)	−	446.3±30.3 (328~557)	338.9±63.7 (354~586)	334.9±34.1 (289.0~370.7)
体宽Body width(BW)	359.75±57.80 (261.00~491.00)	−	322.6±26.0 (229~449)	354.5±60.1 (283~495)	257.6±33.5 (201.0~290.8)
阴门裂长Vulva slit length	33.08±3.70 (26.03~41.65)	36 (30~43)	37.5±2.8 (26~46)	40.3±9.1 (30.3~55.5)	37.0±2.0 (35.0~39.0)
下桥长Underbridge length	75.32±7.90 (61.61~90.40)	78.3 (75~90)	72.0 ±2.9 (65~78)	83.4±8.2 (70~95)	77.5±2.5 (75.0~80.0)
膜孔长Fenestral length	30.82±6.70 (21.82~43.58)	28.5 (25~37)	29.3±2.0 (23~36)	37.4±5.0 (30~50)	40.0±2.0 (38.0~42.0)
膜孔宽Fenestral width	26.80±2.60 (22.65~30.55)	29.8 (25~37)	32.0±2.5 (26-39)	35.1±7.1 (25~47.5)	36.5±0.5 (36.0~37.0)
肛阴距Vulva-anus distance	33.12±5.70 (22.01~41.88)	39.4 (30~46)	−	40.3±9.1 (30.3~55.5)	36.0±1.0 (25.0~37.0)
二龄幼虫(测量标本数)J2(n)	15	25	20	20	10
体长Body length(L)	443.30±32.40 (412.65~468.90)	367 (330~405)	402 ±10.0 (377~450)	461.6±34.5 (404~525)	428.7±8.5 (418~439.4)
口针长Stylet length(S)	20.75±0.80 (18.91~21.60)	18.6 (18.0~19.5)	19.2±0.8 (16~21)	22.5±1.1 (20~25)	18.5±0.9 (17.8~19.1)
唇高Labial region height	3.61±0.40 (3.08~4.50)	3.3 (3.0~3.6)	−	−	3.3±0.4 (3.0~3.5)
唇宽Labial region width	8.57±0.40 (7.71~9.29)	7.7 (7.2~7.8)	−	−	7.8±0.2 (7.6~8.0)
体前端到中食道球距离 Anterior end to median bulb valve	63.66±4.60 (54.00~69.70)	56.4 (52~60)	−	−	69.0±4.3 (65.9~72.0)
尾长Tail length	61.76±5.50 (50.77~72.21)	52.7 (44~57)	57.8±2.9 (47~70)	67.3±6.9 (60~87.5)	65.2±3.1 (63.0~67.4)
透明尾长Tail hyaline region	35.39±3.00 (30.50~40.63)	31.4 (26.0~36.0)	−	37.5±6.4 (30~50)	39.8±7.4 (34.5~45.0)

形态特征 Morphological character	广东清远 Qingyuan, Guangdong, Province, China(This study)	日本 Japan(1974)^[26]	日本 Japan (1992) ^[27]	湖南 Hunan, China (2012) ^[6]	广西 Guangxi, China (2014)^[5]
孢囊(测量标本数)cysts(n)	20	10	50	20	5
体长(不含颈)Body length(excl. neck) (L)	484.90±41.80 (386.00~542.00)	−	446.3±30.3 (328~557)	338.9±63.7 (354~586)	334.9±34.1 (289.0~370.7)
体宽Body width(BW)	359.75±57.80 (261.00~491.00)	−	322.6±26.0 (229~449)	354.5±60.1 (283~495)	257.6±33.5 (201.0~290.8)
阴门裂长Vulva slit length	33.08±3.70 (26.03~41.65)	36 (30~43)	37.5±2.8 (26~46)	40.3±9.1 (30.3~55.5)	37.0±2.0 (35.0~39.0)
下桥长Underbridge length	75.32±7.90 (61.61~90.40)	78.3 (75~90)	72.0 ±2.9 (65~78)	83.4±8.2 (70~95)	77.5±2.5 (75.0~80.0)
膜孔长Fenestral length	30.82±6.70 (21.82~43.58)	28.5 (25~37)	29.3±2.0 (23~36)	37.4±5.0 (30~50)	40.0±2.0 (38.0~42.0)
膜孔宽Fenestral width	26.80±2.60 (22.65~30.55)	29.8 (25~37)	32.0±2.5 (26-39)	35.1±7.1 (25~47.5)	36.5±0.5 (36.0~37.0)
肛阴距Vulva-anus distance	33.12±5.70 (22.01~41.88)	39.4 (30~46)	−	40.3±9.1 (30.3~55.5)	36.0±1.0 (25.0~37.0)
二龄幼虫(测量标本数)J2(n)	15	25	20	20	10
体长Body length(L)	443.30±32.40 (412.65~468.90)	367 (330~405)	402 ±10.0 (377~450)	461.6±34.5 (404~525)	428.7±8.5 (418~439.4)
口针长Stylet length(S)	20.75±0.80 (18.91~21.60)	18.6 (18.0~19.5)	19.2±0.8 (16~21)	22.5±1.1 (20~25)	18.5±0.9 (17.8~19.1)
唇高Labial region height	3.61±0.40 (3.08~4.50)	3.3 (3.0~3.6)	−	−	3.3±0.4 (3.0~3.5)
唇宽Labial region width	8.57±0.40 (7.71~9.29)	7.7 (7.2~7.8)	−	−	7.8±0.2 (7.6~8.0)
体前端到中食道球距离 Anterior end to median bulb valve	63.66±4.60 (54.00~69.70)	56.4 (52~60)	−	−	69.0±4.3 (65.9~72.0)
尾长Tail length	61.76±5.50 (50.77~72.21)	52.7 (44~57)	57.8±2.9 (47~70)	67.3±6.9 (60~87.5)	65.2±3.1 (63.0~67.4)
透明尾长Tail hyaline region	35.39±3.00 (30.50~40.63)	31.4 (26.0~36.0)	−	37.5±6.4 (30~50)	39.8±7.4 (34.5~45.0)