喜旱莲子草与乌蔹莓等植物提取物对松材线虫的毒杀活性研究

doi:10.12302/j.issn.1000-2006.202009032

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南京林业大学学报（自然科学版） ›› 2022, Vol. 46 ›› Issue (4): 45-52.doi: 10.12302/j.issn.1000-2006.202009032

所属专题：松材线虫病致病机理与病害防治

• 专题报道Ⅰ：松材线虫病致病机理与病害防治（执行主编　叶建仁） • 上一篇下一篇

喜旱莲子草与乌蔹莓等植物提取物对松材线虫的毒杀活性研究

江奕(), 陈凤毛()

南京林业大学,南方现代林业协同创新中心,南京林业大学林学院,江苏南京 210037

收稿日期:2020-09-15 修回日期:2021-02-25 出版日期:2022-07-30 发布日期:2022-08-01
通讯作者: 陈凤毛
基金资助:
国家重点研发计划(2017YFD0600104)

Nematocidal activity of extracts of Alternanthera philoxeroides and Cayratia japonica against Bursaphelenchus xylophilus

JIANG Yi(), CHEN Fengmao()

Co-Innovation Center for Sustainable Forestry in Southern China,College of Forestry, Nanjing Forestry University,Nanjing 210037,China

Received:2020-09-15 Revised:2021-02-25 Online:2022-07-30 Published:2022-08-01
Contact: CHEN Fengmao

摘要/Abstract

摘要：

【目的】松材线虫病是当前我国林业生产上危害最严重的森林病害。尽管投入了大量的人力、物力和财力,但松材线虫病防控形势依然严峻。化学杀线剂具有环境局限性,研发高效植物源杀灭松材线虫的药剂对控制松材线虫病疫情,同时降低环境化学污染具有重要意义。【方法】采用乙醇浸渍提取法制备喜旱莲子草(Alternanthera philoxeroides)、乌蔹莓(Cayratia japonica)、迷迭香(Rosmarinus officinalis)等30种植物的提取物,测定其对松材线虫的毒杀活性。利用液液萃取法,用乙醇、石油醚、乙酸乙酯、正丁醇和水等有机溶剂对两种植物中的活性成分进行分步提取,得到药液的质量浓度梯度为10.000、5.000、2.500、1.250和0.625 g/L,并分别测定其对松材线虫的毒杀活性。虫卵悬浮液在25 ℃恒温保湿培养箱内培养,24 h后开始观察,32 h后结束观察。通过镜检统计线虫卵数,并计算相应的线虫卵累计孵化率和孵化抑制率。【结果】采用质量浓度为10 g/L的喜旱莲子草、乌蔹莓乙醇提取物分别处理松材线虫,72 h后线虫校正死亡率均为 100%。喜旱莲子草的乙酸乙酯萃取物和乌蔹莓的水层萃取物中存在主要的杀线活性物质,两种植物萃取物处理松材线虫72 h后的半致死剂量(LC₅₀)分别为0.588 8和0.969 3 g/L。在离体虫卵孵化试验中,喜旱莲子草的乙酸乙酯萃取物对线虫的卵孵化抑制率最大,为79.77%,其次为乌蔹莓的水层萃取物,为46.51%。【结论】可见,喜旱莲子草和乌蔹莓对松材线虫成虫和虫卵均具有较强的生物毒杀活性,可作为新型植物杀线虫剂加以开发利用。

关键词: 喜旱莲子草, 乌蔹莓, 提取物, 松材线虫, 毒杀活性

Abstract:

【Objective】The pine wilt disease is the most serious forest disease in China. Despite considerable investment in manpower, material and financial resources, the prevention and control situation of the pine wilt disease continues to be a serious issue. Due to the limitations of the large-scale application of chemical nematicides, developing high efficiency botanical nematicides to control the pine wilt disease epidemic will be of great importance. 【Method】 The extracts of 30 plants including Alternanthera philoxeroides, Cayratia japonica and Rosmarinus officinalis were prepared using the ethanol impregnation method; their poisonous activity to the pine wilt disease was then determined. Petroleum ether, ethyl acetate, n-butanol and water were used to extract the active ingredients from two plants via the liquid-liquid extraction method. The concentrations of the solutions were 10.000, 5.000, 2.500, 1.250 and 0.625 g/L, respectively, and insecticidal activities against pine wood nematodes (Bursaphelenchus xylophilus) were determined. The egg suspension was cultured in a constant temperature and moisture incubator at 25 ℃, and observation began after 24 h and ended after 32 h. Microscopic examination was used to count the number of nematode eggs, and the corresponding cumulative hatching and hatching inhibition rates of nematode eggs were calculated. 【Result】 (1) The pine wood nematode was treated with ethanol extracts of A. philoxeroides and C. japonica at a mass concentration of 10 g/L; after 72 h, the corrected mortality of nematodes was 100%. (2) The ethyl acetate extract of A. philoxeroides and the aqueous extract of C. japonica were the key nematocidal substances. The Lethal Concentration 50 (LC₅₀) of pine wood nematodes was 0.588 8 and 0.969 3 g/L after 72 h of the treatment. (3) The in vitro egg incubation test demonstrated that the extract from A. philoxeroides in ethyl acetate exhibited the greatest rate of nematode incubation inhibition of 79.77%, followed by the aqueous extract from C. japonica which had an incubation inhibition rate of 46.51%. 【Conclusion】 A. philoxeroides and C. japonica demonstrated the strong inhibitory biological activity for nematode adults and eggs, which may be developed and utilized as new nematicides for plants.

Key words: Alternanthera philoxeroides, Cayratia japonica, extract, Bursaphelenchus xylophilus, toxic activity

中图分类号:

S763.1

江奕,陈凤毛. 喜旱莲子草与乌蔹莓等植物提取物对松材线虫的毒杀活性研究[J]. 南京林业大学学报（自然科学版）, 2022, 46(4): 45-52.

JIANG Yi, CHEN Fengmao. Nematocidal activity of extracts of Alternanthera philoxeroides and Cayratia japonica against Bursaphelenchus xylophilus[J].Journal of Nanjing Forestry University (Natural Science Edition), 2022, 46(4): 45-52.DOI: 10.12302/j.issn.1000-2006.202009032.

图/表 6

表1

供试植物名录"

植物名称 plant name	科名 familia	供试部位 test site	植物名称 plant name	科名 familia	供试部位 test site
喜旱莲子草 Alternanthera philoxeroides	苋科 Amaranthaceae	全株 whole plant	蛇床 Cnidium monnieri	伞形科 Apiaceae	全株 whole plant
乌蔹莓 Cayratia japonica	葡萄科 Vitaceae	全株 whole plant	常春藤 Hedera nepalensis	五加科 Araliaceae	茎叶 stem and leaf
菖蒲 Acorus calamus	天南星科 Araceae	叶 leaf	大戟 Euphorbia pekinensis	大戟科 Euphorbiaceae	全株 whole plant
迎春 Jasminum nudiflorum	木樨科 Oleaceae	叶 leaf	万年青 Rohdea japonica	天门冬科 Asparagaceae	全株 whole plant
络石 Trachelospermum jasminoides	夹竹桃科 Apocynaceae	茎叶 stem and leaf	龙葵 Solanum nigrum	茄科 Solanaceae	全株 whole plant
泽漆 Euphorbia helioscopia	大戟科 Euphorbiaceae	全株 whole plant	紫花地丁 Viola philippica	堇菜科 Violaceae	全株 whole plant
玉簪 Hosta plantaginea	百合科 Liliaceae	叶 leaf	千里光 Senecio scandens	菊科 Asteraceae	茎叶 stem and leaf
侧柏 Platycladus orientalis	柏科 Cupressaceae	叶 leaf	荆芥 Nepeta cataria	唇形科 Lamiaceae	全株 whole plant
大花旋复花 Inula britanica	菊科 Asteraceae	花 flower	野豌豆 Vicia sepium	豆科 Leguminosae	全株 whole plant
辣蓼 Polygonum flaccidum	蓼科 Polygonaceae	全株 whole plant	商陆 Phytolacca acinosa	商陆科 Phytolaccaceae	全株 whole plant
冬青卫矛 Euonymus japonicus	卫矛科 Celastraceae	叶 leaf	粉花绣线菊 Spiraea japonica	蔷薇科 Rosaceae	全株 whole plant
鸢尾 Iris tectorum	鸢尾科 Iridaceae	全株 whole plant	酢浆草 Oxalis corniculata	酢浆草科 Oxalidaceae	全株 whole plant
飞蓬 Erigeron acer	菊科 Asteraceae	全株 whole plant	迷迭香 Rosmarinus officinalis	唇形科 Lamiaceae	茎叶 stem and leaf
金丝桃 Hypericum monogynum	藤黄科 Guttiferae	花叶 flowers and leaves	荚蒾 Viburnum dilatatum	忍冬科 Caprifoliaceae	枝叶 branches and leaves
老鹳草 Geranium wilfordii	牦牛儿苗科 Geraniaceae	全株 whole plant	地锦 Parthenocissus tricuspidata	葡萄科 Vitaceae	茎叶 stem and leaf

表1

表2

植物提取物对松材线虫的毒杀活性"

供试植物 tested plant	校正死亡率/% corrected mortality (mean±SD)		活性强度 activity degree
供试植物 tested plant	24 h	48 h	24 h	48 h
喜旱莲子草 Alternanthera philoxeroides	100.00±0.00	100.00±0.00	++++	++++
乌蔹莓 Cayratia japonica	100.00±0.00	100.00±0.00	++++	++++
菖蒲 Acorus calamus	13.88±1.92	17.59±3.24	+	+
迎春 Jasminum nudiflorum	21.51±7.13	23.47±2.44	+	+
络石 Trachelospermum jasminoides	28.65±3.73	42.05±4.21	+	++
泽漆 Euphorbia helioscopia	32.84±5.56	54.56±3.77	++	+++
玉簪 Hosta plantaginea	20.49±0.96	23.93±2.26	+	+
侧柏 Platycladus orientalis	13.25±6.53	21.23±8.47	+	+
大花旋复花 Inula britanica	14.89±2.74	34.45±3.37	+	++
辣蓼 Polygonum flaccidum	37.97±4.07	51.84±4.11	++	+++
冬青卫矛 Euonymus japonicus	28.46±3.52	39.07±2.59	+	++
鸢尾 Iris tectorum	28.25±3.14	28.93±2.73	+	+
飞蓬 Erigeron acer	26.59±2.18	29.40±2.11	+	+
金丝桃 Hypericum monogynum	7.96±1.12	19.52±2.09	-	+
老鹳草 Geranium wilfordii	29.46±5.50	34.94±4.33	++	++
蛇床 Cnidium monnieri	11.51±2.91	13.05±2.13	+	+
常春藤 Hedera nepalensis	11.24±1.21	21.89±2.56	+	+
大戟 Euphorbia pekinensis	33.83±3.11	47.72±4.23	++	++
万年青 Rohdea japonica	12.64±0.69	15.85±2.77	+	+
龙葵 Solanum nigrum	39.27±4.56	45.11±5.58	++	++
紫花地丁 Viola philippica	59.56±4.01	63.47±5.37	+++	+++
千里光 Senecio scandens	19.08±2.59	34.62±2.78	+	++
荆芥 Nepeta cataria	31.65±2.02	36.85±3.13	++	++
野豌豆 Vicia sepium	30.34±3. 83	43.83±7.47	++	++
商陆 Phytolacca acinosa	24.81±3.55	27.66±1.16	+	+
粉花绣线菊 Spiraea japonica	8.57±1.11	13.24±1.24	-	+
酢浆草 Oxalis corniculata	25.84±0.63	37.78±2.09	+	++
迷迭香 Rosmarinus officinalis	57.52±5.12	72.15±7.52	+++	+++
荚蒾 Viburnum dilatatum	17.41±1.14	29.35±3.55	+	+
地锦 Parthenocissus tricuspidata	15.85±0.86	18.44±2.07	+	+
溶剂对照	0.00±0.00	0.00±0.00

表2

表3

图1

表4

表5

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处理液 treatment solution	质量浓度/ (g·L^-1) mass concentration	校正死亡率/% corrected mortality
处理液 treatment solution	质量浓度/ (g·L^-1) mass concentration	24 h	48 h	72 h
喜旱莲子草乙酸乙酯萃取物 ethyl acetate extract of Alternanthera philoxeroides	10.000	100.00±0.00 a	100.00±0.00 a	100.00±0.00 a
	5.000	94.91±2.71 a	96.76±2.32 a	100.00±0.00 a
	2.500	78.41±4.33 b	86.92±4.11 b	93.54±3.09 b
	1.250	53.64±4.62 c	74.20±4.29 c	86.26±2.24 c
	0.625	35.86±3.52 d	51.50±2.86 d	73.06±4.67 d
乌蔹莓水层萃取物 aqueous extract of Cayratia japonica	10.000	86.45±1.49 a	97.53±1.11 a	100.00±0.00 a
	5.000	79.29±5.28 b	93.17±3.88 a	98.91±0.99 a
	2.500	68.04±2.57 c	74.25±1.45 b	83.23±4.39b
	1.250	27.10±3.55 d	31.57±4.72 c	40.81±3.59 c
	0.625	8.38±1.80 e	10.97±4.60 d	17.34±2.07 d
溶剂对照 solvent control		0.00±0.00	0.00±0.00	0.00±0.00

处理 treatment	累计孵化率/% egg hatching rate			孵化抑制率/% control rate of hatching
处理 treatment	24 h	28 h	32 h	孵化抑制率/% control rate of hatching
喜旱莲子草乙酸乙酯萃取物 ethyl acetate extract of Alternanthera philoxeroides	15.43±0.29	18.69±0.52	19.08±0.96	79.77
乌蔹莓水层萃取物 aqueous extract of Cayratia japonica	29.38±0.74	43.90±0.48	50.45±0.62	46.51
溶剂对照 solvent control	46.74±1.49	80.23±1.71	94.31±0.57

喜旱莲子草与乌蔹莓等植物提取物对松材线虫的毒杀活性研究

Nematocidal activity of extracts of Alternanthera philoxeroides and Cayratia japonica against Bursaphelenchus xylophilus

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供试植物 tested plant	质量浓度/ (g·L^-1) mass concentration	72 h校正死亡率/% 72 h corrected mortality	活性强度 activity degree
喜旱莲子草 Alternanthera philoxeroides	10.000	100.00±0.00 a	++++
	5.000	100.00±0.00 a	++++
	2.500	89.65±2.93 b	++++
	1.250	65.45±6.00 c	+++
	0.625	40.91±2.11 d	++
乌蔹莓 Cayratia japonica	10.000	100.00±0.00 a	++++
	5.000	100.00±0.00 a	++++
	2.500	87.32±5.85 b	++++
	1.250	47.11±3.87 c	++
	0.625	24.62±2.69 d	+

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