香榧藻斑病病原的分离鉴定及防治药剂筛选

doi:10.11707/j.1001-7488.LYKX20220537

摘要/Abstract

摘要：

目的: 探明香榧藻斑病的病原种类及其生物学特性，筛选对香榧藻斑病具有良好防治效果的药剂，为香榧藻斑病防治提供科学依据。方法: 在浙江省香榧主产区采集藻斑病病叶，结合传统形态学方法与分子生物学手段对从病叶中分离获得的香榧绿藻进行鉴定，明确病原藻的生物学特性，通过室内毒力测定和林间防治试验筛选防治香榧藻斑病的优良药剂。结果: 1）从香榧藻斑病病叶中分离获得3种病原绿藻类群，结合藻株形态学特点以及SSU-ITS区间序列分析，3种绿藻类群分别被鉴定为链带藻、软克里藻和Tritostichococcus corticulus，均为香榧病原新记录种。3种香榧绿藻的培养条件相似，均适宜在初始pH 11、温度25 ℃、光照/黑暗为12 h/12 h光周期下的BG-11培养基中生长。2）以混合藻为靶标进行室内毒力测定试验显示，12%松脂酸铜悬浮剂灭藻效果最好，EC₅₀可达8.33 mg·L^?1，显著低于其他药剂（P<0.05），其后依次为33.5%喹啉铜悬浮剂、80%波尔多液、20%噻菌铜悬浮剂和45%石硫合剂。林间防治试验表明，施药30天后，12%松脂酸铜和33.5%喹啉铜悬浮剂的防治效果分别达78.11%和71.62%。结论: 从浙江省香榧上新发现3种病原绿藻类群，藻群生物学特性类似。室内毒力测定和林间防治试验表明，12%松脂酸铜和33.5%喹啉铜悬浮剂对香榧藻斑病有较好防治效果。

关键词: 香榧, 藻斑病, 鉴定, 生物学特性, 毒力测定, 林间防治

Abstract:

Objective: The study aims to explore the pathogens of algal spots of Torreya grandis cv. ‘Merrillii’ and their biological characteristics, as well as screen efficient algicides, so as to provide scientific basis for controlling the disease. Method: The diseased leaves of T. grandis cv. ‘Merrillii’ were collected from the main planting areas in Zhejiang Province, and the algae were isolated from diseased leaves, and then identified by traditional morphological observation combined with molecular biological methods. The biological characteristics of the pathogenic algae were studied, and efficient algicides for controlling the disease were screened by laboratory virulence assay and the field control experiment. Result: 1) Three groups of synantectic pathogenic algae were isolated from algal spots, and they were identified as Desmodesmus armatus, Klebsormidium flaccidum and Tritostichococcus corticulus, respectively, based on the morphological characteristics and the SSU-ITS sequence analysis, all of which were newly reported pathogens of T. grandis cv. ‘Merrillii’. The culture conditions of the three alga species were similar, and they all grew well in BG-11 medium at 25 ℃ with an initial pH of 11 and a photoperiod of 12 h light/12 h dark. 2) With the mixed algae as the target, a laboratory toxicity assay showed that 12% copper rosinate suspension had the best algal killing effect, and the EC₅₀ was 8.33 mg·L^?1, which was significantly lower than other algicides （P<0.05）, followed by 33.5% copper quinoline suspension agent, 80% Bordeaux mixture, 20% thiophanate copper suspension agent and 45% lime sulfur mixture. The field control test showed that after 30 days of application, the control effects of 12% copper rosinate and 33.5% copper quinoline suspending agent were 78.11% and 71.62%, respectively. Conclusion: Three pathogenic green algae have newly been discovered from T. grandis cv. ‘Merrillii’ in Zhejiang Province, and the biological characteristics of the three alga species are similar. Laboratory toxicity assay and field control test show that 12% copper rosinate and 33.5% copper quinoline suspending agent have good control effect on algal spot of T. grandis cv. ‘Merrillii’.

Key words: Torreya grandis cv. ‘Merrillii’, algal spot, identification, biological characteristics, toxicity test, field control

中图分类号:

刘艾涛,叶碧欢,陈友吾,宋其岩,李海波,沈建军,张昕. 香榧藻斑病病原的分离鉴定及防治药剂筛选[J]. 林业科学, 2024, 60(1): 111-119.

Aitao Liu,Bihuan Ye,Youwu Chen,Qiyan Song,Haibo Li,Jianjun Shen,Xin Zhang. Isolation and Identification of Pathogens of Algal Spots of Torreya grandis cv. ‘Merrillii’ and Screening of the Algicides[J]. Scientia Silvae Sinicae, 2024, 60(1): 111-119.

图/表 10

图1

图2

图3

图4

图5

图6

图7

图8

表1

表2

参考文献 0

	戴文圣, 黎章矩, 程晓建, 等. 杭州市香榧生产的发展前景与对策. 浙江林学院学报, 2006, (3): 334- 337.
	Dai W S, Li Z J, Cheng X J, et al. Development future and strategies of production of Torreya grandis‘Merrillii’ in Hangzhou. Journal of Zhejiang Forestry College, 2006, (3): 334- 337.
	何美仙, 黄飞来, 刘忠良. 山茶叶部主要病害的发生与防治. 现代农业科技, 2017, (14): 110- 111. doi: 10.3969/j.issn.1007-5739.2017.14.073
	He M X, Huang F L, Liu Z L. Occurrence and control of main diseases in Camellia japonica leaves. Modern Agricultural Science and Technology, 2017, (14): 110- 111. doi: 10.3969/j.issn.1007-5739.2017.14.073
	吕军美, 郑文彪, 刘金龙, 等. 2015. 石硫合剂防治香榧绿藻的试验. 南方林业科学, 43(3): 30–32, 54.
	Lv J M, Zheng W B, Liu J L, et al. 2015. Test of LSSS control of Chlorella. South China Forestry Science, 43(3): 30–32 , 54. ［in Chinese］
	欧阳峥嵘, 温小斌, 耿亚红, 等. 光照强度、温度、pH、盐度对小球藻(Chlorella)光合作用的影响. 武汉植物学研究, 2010, 28 (1): 49- 55.
	Ouyang Z R, Wen X B, Geng Y H, et al. The effects of light intensities, temperatures, pH and salinities on photosynthesis of Chlorella. Journal of Wuhan Botanical Research, 2010, 28 (1): 49- 55.
	王大平, 李道高. 夏橙绿斑病的病原鉴定及其生长特性研究. 中国农学通报, 2005, 21 (6): 288- 291. doi: 10.3969/j.issn.1000-6850.2005.06.087
	Wang D P, Li D G. Studies on the identification and growth characteristics of the pathogen of Valencia orange green spot. Chinese Agricultural Science Bulletin, 2005, 21 (6): 288- 291. doi: 10.3969/j.issn.1000-6850.2005.06.087
	王　鸿, 郭　涛, 应国清. 榧属植物活性成分与药理作用研究进展. 中草药, 2007, 38 (11): 1747- 1750. doi: 10.3321/j.issn:0253-2670.2007.11.052
	Wang H, Guo T, Ying G Q. Advances in studies on active constituents and their pharmacological activities for plants of Torreya Arn. Chinese Traditional and Herbal Drugs, 2007, 38 (11): 1747- 1750. doi: 10.3321/j.issn:0253-2670.2007.11.052
	王　霞, 吕宪国, 张学林, 等. 松花湖铜绿微囊藻无菌株和单藻株生长因素的研究. 生态学杂志, 2005, 24 (5): 518- 522. doi: 10.3321/j.issn:1000-4890.2005.05.011
	Wang X, Lü X G, Zhang X L, et al. Growth factors of unialgal strain and axenic strain of Microcystis aeruginosa in Songhua Lake. Chinese Journal of Ecology, 2005, 24 (5): 518- 522. doi: 10.3321/j.issn:1000-4890.2005.05.011
	吴连海, 吴黎明, 倪荣新, 等. 香榧栽培经济效益分析. 浙江农林大学学报, 2013, 30 (2): 299- 303. doi: 10.11833/j.issn.2095-0756.2013.02.023
	Wu L H, Wu L M, Ni R X, et al. Economic benefits of Torreya grandi‘Merrillii’ plantings. Journal of Zhejiang A & F University, 2013, 30 (2): 299- 303. doi: 10.11833/j.issn.2095-0756.2013.02.023
	许　珍. 2017. 温度和光强对四种藻类生理生态学特性的影响研究. 武汉: 长江科学院.
	Xu Z. 2017. Effects of temperature and light intensity on physiological and ecological characteristics of four algae. Changjiang Academy of Sciences. ［in Chinese］
	徐志宏, 吾中良, 陈秀龙, 等. 浙江省香榧病虫害及害虫天敌种类调查. 中国森林病虫, 2005, 24 (1): 14- 19. doi: 10.3969/j.issn.1671-0886.2005.01.006
	Xu Z H, Wu Z L, Chen X L, et al. Investigation on insect pests and diseases in Torreya grandis and their natural enemies in Zhejiang Province. Forest Pest and Disease, 2005, 24 (1): 14- 19. doi: 10.3969/j.issn.1671-0886.2005.01.006
	叶晓明, 钱宇汀, 叶　雯, 等. 香榧绿藻的生物学特性及物种鉴定. 浙江农林大学学报, 2019, 36 (4): 629- 637. doi: 10.11833/j.issn.2095-0756.2019.04.001
	Ye X M, Qian Y T, Ye W, et al. Biological characteristics and species identification of Chlorella sp. with Torreya grandis ‘Merrillii’. Journal of Zhejiang A & F University, 2019, 36 (4): 629- 637. doi: 10.11833/j.issn.2095-0756.2019.04.001
	郑宝荣, 叶建仁, 王记祥, 等. 肉桂藻斑病的初步研究. 中国森林病虫, 2004, 23 (2): 8- 10. doi: 10.3969/j.issn.1671-0886.2004.02.003
	Zheng B R, Ye J R, Wang J X, et al. Algae-spot disease in Cinnamomum cassia. Forest Pest and Disease, 2004, 23 (2): 8- 10. doi: 10.3969/j.issn.1671-0886.2004.02.003
	Browne F B, Brannen P M, Scherm H, et al. 2019. Evaluation of disinfectants, algicides, and fungicides for control of orange cane blotch of blackberry in the field. Crop Protection, 112–117.
	Cai Z P, Duan S S, Zhu H H. Optical density method and cell count method for determining the growth of three energy microalgae and their correlation and verification. Journal of Southern Agriculture, 2012, 43 (10): 1480- 1484.
	Chandel N, Ahuja V, Gurav R, et al. Progress in microalgal mediated bioremediation systems for the removal of antibiotics and pharmaceuticals from wastewater. Science of the Total Environment, 2022, 825, 153895. doi: 10.1016/j.scitotenv.2022.153895
	Chanthapatchot W, Satjarak A. Distribution, diversity, and specificity of a parasitic algal genus Cephaleuros in Thailand. Sains Malaysiana, 2019, 48 (8): 1609- 1618. doi: 10.17576/jsm-2019-4808-06
	Glaser K, Donner A, Albrecht M, et al. Habitat-specific composition of morphotypes with low genetic diversity in the green algal genus Klebsormidium (Streptophyta) isolated from biological soil crusts in Central European grasslands and forests. European Journal of Phycology, 2017, 52 (2): 188- 199. doi: 10.1080/09670262.2016.1235730
	Han K S, Park M J, Park J H, et al. First report of algal leaf spot associated with Cephaleuros virescens on greenhouse-grown Ficus benghalensis in Korea. Australasian Plant Disease Notes, 2011, 6 (1): 72- 73. doi: 10.1007/s13314-011-0024-8
	Karsten U, Herburger K, Holzinger A. Living in biological soil crust communities of African Deserts—physiological traits of green algal Klebsormidium species (Streptophyta) to cope with desiccation, light and temperature gradients. Journal of Plant Physiology, 2016, 194, 2- 12. doi: 10.1016/j.jplph.2015.09.002
	Li L, Cui J, Liu Q, et al. Screening and phylogenetic analysis of lipid-rich microalgae. Algal Research, 2015, 11, 381- 386. doi: 10.1016/j.algal.2015.02.028
	Lin C S, Lin Y H, Wu J T. Biodiversity of the epiphyllous algae in a Chamaecyparis forest of northern Taiwan. Botanical Studies, 2012, 53, 489- 499.
	Marin B, Palm A, Klingberg M, et al. Phylogeny and taxonomic revision of plastid-containing euglenophytes based on SSU rDNA sequence comparisons and synapomorphic signatures in the SSU rRNA secondary structure. Protist, 2003, 154 (1): 99- 145. doi: 10.1078/143446103764928521
	öztürk B Y. 2019. Intracellular and extracellular green synthesis of silver nanoparticles using Desmodesmus sp.: their Antibacterial and antifungal effects. Caryologia. International Journal of Cytology, Cytosystematics and Cytogenetics, 72(1): 29–43.
	Pitaloka M K, Petcharat V, Arikit S, et al. Cephaleuros virescens, the cause of an algal leaf spot on Para rubber in Thailand. Australasian Plant Disease Notes, 2015, 10 (1): 1- 4. doi: 10.1007/s13314-014-0152-z
	Pröschold T, Darienko T. The green puzzle Stichococcus (Trebouxiophyceae, Chlorophyta): new generic and species concept among this widely distributed genus. Phytotaxa, 2020, 441 (2): 113- 142. doi: 10.11646/phytotaxa.441.2.2
	Ryšánek D, Holzinger A, Škaloud P. Influence of substrate and pH on the diversity of the aeroterrestrial alga Klebsormidium (Klebsormidiales, Streptophyta): a potentially important factor for sympatric speciation. Phycologia, 2016, 55 (4): 347- 358. doi: 10.2216/15-110.1
	Sunpapao A, Thithuan N, Bunjongsiri P, et al. Cephaleuros parasiticus, associated with algal spot disease on Psidium guajava in Thailand. Australasian Plant Disease Notes, 2016a, 11 (1): 1- 4. doi: 10.1007/s13314-015-0186-x
	Sunpapao A. , Thithuan N, Pitaloka M K , et al. 2016b. Algal leaf spot of Lansium parasiticum caused by Cephaleuros sp. in Thailand. Journal of Plant Pathology, 98(2): 369.
	Vasconcelos C V, Teodoro Pereira F, dos Santos Galvão C, et al. Occurrence of algal leaf spot (Cephaleuros virescens Kunze) on avocado in Goiás State, Brazil. Summa Phytopathologica, 2016, 42 (1): 108. doi: 10.1590/0100-5405/2109

药剂 Pesticide	毒力回归方程 Toxic regression equation	有效中浓度 EC₅₀/（mg·L^?1）	R²
12%松脂酸铜悬浮剂 12% copper rosinate suspension	Y = 1.722 x?1.585	8.33a	0.937
33.5%喹啉铜悬浮剂 33.5% copper quinoline suspension	Y = 0.229 x?0.388	49.68b	0.999
80%波尔多液 80% Bordeaux mixture	Y = 5.592 x?11.207	100.96c	0.973
20%噻菌铜剂悬浮剂 20% Copper thiabendate suspension	y = 1.288 x?2.674	119.07c	0.976
45%石硫合剂 45% lime sulfur mixture	Y = 1.352 x?4.658	2778.18d	0.993

处理 Treatment	施药前的病情指数 Condition index before treatment	施药后的病情指数 Condition index after treatment	防治效果 Control efficiency （%）
12%松脂酸铜悬浮剂 12% copper rosinate suspension	64.44	15.00	78.11
33.5%喹啉铜悬浮剂 33.5% copper quinoline suspension	61.67	18.61	71.62
CK	63.33	67.20	—

[1]	杨忠岐,曹亮明,王小艺,刘慧慧,吴耀军,蒋学建,罗缉. 一种重要的桉树外来入侵害虫——桉树叶瘿球角姬小蜂（膜翅目：姬小蜂科）[J]. 林业科学, 2023, 59(8): 85-89.
[2]	张元臣,鲍梦楠,薛爽,王璞,王兴云,张坤朋,王景顺. 汤川瘿蜂寄生性天敌调查及其优势天敌长尾小蜂生物学特性[J]. 林业科学, 2023, 59(6): 112-117.
[3]	沈家怡,吴翠萍,姚佳,吴家胜,张瑞,胡渊渊. 香榧假种皮开裂过程中组织结构、细胞壁代谢的变化[J]. 林业科学, 2023, 59(2): 86-95.
[4]	成豪,吴家胜,马爽,仲嘉玥,胡渊渊,喻卫武,俞晨良,宋丽丽,索金伟. 树形调整对香榧成花和坐果的影响[J]. 林业科学, 2023, 59(11): 49-58.
[5]	李俊楠,朱禹臻,陈润恺,付煜,蔡梦玲,黄炳荣,徐云,吴松青,张飞萍. 松天牛小首螨的研究Ⅰ.生物学特性[J]. 林业科学, 2022, 58(6): 79-87.
[6]	郑刘辉,侯宇,张新凤,喻卫武,曾燕如,戴文圣. 香榧种子生长发育过程中假种皮挥发油的变化[J]. 林业科学, 2022, 58(11): 127-136.
[7]	彭欣,王瀚棠,郭春晖,杨振德,周静,王雪,丁芷柔. 入侵性致瘿害虫桉树枝瘿姬小蜂(膜翅目: 姬小蜂科)EST-SSR开发及隐种鉴定[J]. 林业科学, 2021, 57(9): 140-151.
[8]	陈越渠,刘庆珍,李立梅,张杨,韩姣,张永安. 杨树溃疡病拮抗链霉菌的筛选及鉴定[J]. 林业科学, 2021, 57(7): 92-100.
[9]	任利利,陶静,武海卫,宗世祥,王传珍,华德,石娟,刘漪舟,骆有庆. 重大害虫长林小蠹入侵我国的首次发现与侵染特征[J]. 林业科学, 2021, 57(5): 140-150.
[10]	万琼莲,王连春,王泉,许杏萍,苏帆,赵静,蔡红. 羽脉山黄麻丛枝植原体的分子鉴定及病害调查[J]. 林业科学, 2021, 57(5): 195-201.
[11]	杨雨桐,张卿硕,符韵林,孙静. 巴里黄檀心材色素成分及其稳定性和抗氧化性[J]. 林业科学, 2021, 57(3): 126-134.
[12]	韦宜慧,陈嘉琪,赵光宇,董玉红,厚凌宇,焦如珍. 杉木林土壤和苗木内生溶磷细菌的筛选及其溶磷特性[J]. 林业科学, 2020, 56(12): 1-9.
[13]	贺然,崔夏,应玥,曲良建,王瑞珍,张永安. 防治松褐天牛成虫的白僵菌菌株室内筛选和鉴定[J]. 林业科学, 2020, 56(10): 129-134.
[14]	张亚波,张威,张守科,彭瀚,孟珂,舒金平,王浩杰. 4株绿僵菌的鉴定及对松褐天牛成虫致病效果比较[J]. 林业科学, 2020, 56(1): 103-111.
[15]	马涛, 黄志嘉, 朱映, 林娜, 肖强, 何余容, 温秀军. 尺蛾科昆虫性信息素组分特征及应用进展[J]. 林业科学, 2019, 55(5): 152-162.