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研究生:胡博隆
研究生(外文):Bo-Long Hu
論文名稱:探討藥用植物百部萃取物對爪哇根瘤線蟲之防治效果
論文名稱(外文):Nematicidal potency of Stemona spp. (Stemonaceae) herb’s extracts against root-knot nematode, Meloidogyne javanica (Treub) Chitwood.
指導教授:楊瓊儒
指導教授(外文):Chiung-Ru Yang
學位類別:碩士
校院名稱:國立嘉義大學
系所名稱:生物資源學系研究所
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2006
畢業學年度:94
語文別:中文
論文頁數:98
中文關鍵詞:爪哇根瘤線蟲百部防治
外文關鍵詞:Meloidogyne javanica (Treub) ChitwoodStemona spp.control
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百部為百部科(Stemonaceae)百部屬(Stemona spp.)之植物,因其根部含有多種生物鹼具有毒殺甜菜夜蛾、小菜蛾與牛蜱的能力,已被引用為天然殺蟲化學物之研究。本文由百部、杏仁、銀杏、油松節、連翹、黑丑、馬鞭草、槐角、側柏葉、扁蓄、黃柏及烏梅等12種中藥材篩選出對葉百部(Stemona tuberosa Lour.)及市售百部(Stemonae Radix)的塊根在20%水煮液下對爪哇根瘤線蟲(Meloidogyne javanica (Treub) Chitwood)二齡幼蟲致死率分別高達64.5%及70.5%。對葉百部採自嘉義縣石硦林場,多為新鮮塊根,市售百部則購於中藥店多為乾燥塊根,比較二者水煮液殺線蟲之效果,發現1%及5%水煮液下分別呈5%及1%顯著性差異,10%及20%水煮液則無顯著性差異,但都與對照組呈1%極顯著性差異,尤以20%水煮液最高,因此,為節省經費及方便貯存,除將對葉百部塊根以烘箱乾燥處理外,水煮液採用20%濃度。百部塊根水煮液的線蟲致死率及抑制卵粒孵化的效果皆大於莖部或葉片的水煮液。百部乾燥塊根分別以蒸餾水、甲醇、丙酮萃取,結果發現對葉百部水萃取物10%稀釋濃度、甲醇萃取物15%稀釋濃度及丙酮萃取物5%稀釋濃度,與市售百部水萃取物15%稀釋濃度、甲醇萃取物15%稀釋濃度及丙酮萃取物5%稀釋濃度的防治效果皆佳,故被選用為本文最適稀釋濃度。12種中藥材的pH值對線蟲致死率並無極大的影響(r=-0.234),烏梅水煮液雖為強酸性(pH1.5),但線蟲致死率僅有16%,而百部不同萃取物最適稀釋濃度,經1N NaOH調整pH為7.0,與原有pH值之線蟲致死率皆無顯著性差異。六種百部不同萃取物在-20℃下貯放二個月,也不影響其殺線蟲效果。在番茄TK70栽盆試驗中,六種百部不同萃取物最適稀釋濃度對爪哇根瘤線蟲之根瘤指數(每克根重的根瘤數),以移植前7天施用20ml之效果最佳為2.0~7.3,與對照組之27.0呈1%極顯著性差異。施用時期對根瘤指數、每百克土壤線蟲量及卵塊孵化數上,處理間皆達極顯著性差異,但施用量僅在根瘤指數上,處理間有顯著性差異(p=0.0026)。施用時期與施用量對爪哇根瘤線蟲之防治並無交感效應。在番茄生長方面,根重、植物鮮重及株高以移植前處理者較佳。施用時期對於根重及鮮重上,處理間皆達顯著性差異。施用量僅在根重上,處理間達顯著性差異。施用時期與施用量對番茄之生長無交感效應。六種百部萃取物施用於番茄3週齡移植株時,防治效果較佳。移植株週齡對根瘤指數、每百克土壤線蟲量及卵塊孵化數上,處理間皆達顯著性差異,而施用量僅有根瘤指數在處理間有顯著性差異。移植株週齡與施用量的交感效應只在根瘤指數上有顯著性差異。移植株週齡越高,生長情形越佳,與對照組大多呈極顯著性差異,但為兼顧防治效果,擬建議番茄在3週齡移植株時即可施用百部萃取物。對葉百部甲醇萃取物施用三次,或市售百部甲醇萃取物施用一次,對爪哇根瘤線蟲的防治效果最好,其平均根瘤指數為9.3~9.56與接種線蟲對照組及托福松對照組之17.14~18.02呈極顯著性差異。番茄生長情形則以市售百部水萃取物施用一次者較接近空白對照組及托福松處理,且優於線蟲對照組。本文作者認為台灣的對葉百部具有發展潛力,可使用水萃取物在病土經移植前處理後,等番茄株齡較大時,再施用一次。
The herbs Stemona spp. (Stemonaceae), known as “Bai-Bu” in traditional Chinese Medicine, with tuberous roots have many kinds of alkaloids which proved to kill Spodptera exigua, Plutella xylostella and tick (Boophilus microplus), The effects of Stemona spp. extracts on root-knot nematode, Meloidogyne javanica, were investigated as be natural pesticides. We collected twelve Chinese medicinal herbs including Stemona spp., Prunus armeniaca, Ginkgo biloba, Pinus massoniana, Forsythia suspensa, Pharbitis purpurea, Verbena officinalis, Sophora japonica, Biota orientalis, Polygonum aviculare , Phellodendron chinense and Prunus mume, and found the tubers of Stemonae Radix and Stemona tuberosa Lour. among herbs had the best control on Meloidogyne javanica 2nd-stage juveniles (J2) which showed mortality as 64.5% and 70.5%, respectively, under 20% crude extracts. S. tuberosa with fresh tubers collected from the Chiayi county shi-long farm in Taiwan, while Stemonae Radix with dry tubers is commercial product bought from the traditional Chinese medicine store. Under 1% and 5% crude extracts, there were 5% and 1%, respectively, significantly difference between Stemonae Radix and S. tuberosa on the mortality of M. javanica J2, but not with 10% and 20% crude extracts. All crude extracts showed good control efficiency on root-knot nematode, and 20% crude extract was the best of all, and was chosen to conduct the experiments for the reason of fund and storage. Tuber’s crude extracts were re-extracted with distilled water, methanol or acetone, S. tuberosa extracts were chosen 10%, 15% and 5% dilutions, respectively, and Stemonae Radix extracts were chosen 15%, 15% and 5% dilutions, respectively, as the optimal concentration of control efficiency. There was little relationship (r=-0.234) between the pH value of twelve Chinese medicinal herbs and the mortality of J2. Even if strong acidic as Prunus mume (pH=1.5) only had 16% mortality of J2. Six Stemona spp. extract optimal dilutions, pH value were adjusted to 7.0 by 1N NaOH, and showed no significantly difference with original pH value on the mortality of J2. These extract dilutions storaged under -20℃ for two months didn’t affect the nematicidal efficiency. In the tomato TK70 pot trail, the optimal dilutions of six Stemona spp. extracts 20 ml applicated on 7 days before transplant had 1% significantly difference with Nema CK on root-knot index (galls/g. root). Application date had significantly difference among treatments on root-knot index, nematodes per 100g soil and egg masses hatch, while application dose only had significantly difference (p=0.0026) among treatments on root-knot index. There were no interactions between application date and dose on the control of M. javanica. On root weight, plant weight and plant height of tomato treated on 7 days before transplant were better than on transplant treatment. Application dose had significantly difference among treatments only on root weight, and no interactions between application date and dose on the growth of tomato TK70. Concerning plant age, six Stemona spp. extracts had better control on 3-weeks transplant. Transplant date had significantly difference among treatments on root-knot index, nematodes per 100g soil and egg masses hatch, while application dose only on root-knot index. There was interaction between transplant date and dose on root-knot index. The later transplant, the better growth of tomato. Although transplant date had significantly difference among treatments on the growth of tomato, we suggest that use Stemona spp. extracts on 3-weeks transplant for control efficiency. Apply 3 times of Stemona tuberosa methanol extract or one application of Stemonae Radix methanol extract had the best control efficiency among treatments. The growth of tomato treated by Stemonae Radix water extract one application were the same as Blank CK and Terbofus CK, and better than Nema CK. So, we suggest that Stemona tuberosa is potential on the control of M. javanica, and we can use water extract on 7 days before transplant and again after plant get old.
中文摘要………………………………………………………………1
英文摘要………………………………………………………………4
前言……………………………………………………………………7
前人研究…………………………………………………………….10
材料與方法…………………………………………………….…..24
(一)、供試材料………………………………………………….24
一、供試植物………………………………………...……………24
二、供試土壤………………………………………...……………24
三、供試根瘤線蟲之接種源………………………...……………24
四、供試藥用植物資材……………………………...……………25
五、儀器設備………………………………………...……………25(二)、試驗方法………………………………………………….26
一、供試中藥材20%水煮液之製備…………………...….………26
二、百部不同溶劑萃取物之製備……………………...…………27
1. 供試藥材萃取物對爪哇根瘤線蟲二齡幼蟲的致死率及卵粒孵化率之影響....…………………………….……………………....…27
(1). 供試中藥材20%水煮液對根瘤線蟲二齡幼蟲之致死率…...28
(2). 對葉百部新鮮塊根與乾燥塊根水煮液對根瘤線蟲二齡幼蟲致死率之影響……….…………………….....…….………........28
(3). 百部不同部位的20%水煮液對根瘤線蟲二齡幼蟲致死率及卵粒孵化率之影響….…………………..………………............…28
(4). 百部之不同溶劑萃取物對根瘤線蟲二齡幼蟲致死率及卵粒孵化率之影響………………………………………..........……….29
(5). 百部萃取物的pH值對根瘤線蟲二齡幼蟲致死率之影響....29
(6). 百部不同溶劑萃取物的貯放時間對根瘤線蟲二齡幼蟲致死率之影響…………………………………………………........…...30
2. 百部不同溶劑之萃取物對爪哇根瘤線蟲防治效果與番茄TK70生育之影響………………………..……………………….....………30
(1). 百部不同溶劑之萃取物以不同施用量及不同施用時間對番茄根瘤線蟲之防治效果…………………………………...........…30
(2). 百部不同溶劑之萃取物以不同施用量在不同週齡的番茄移植苗上施用對根瘤線蟲之防治效果…………………….........…..31
(3). 百部不同溶劑之萃取物的施用次數對持效性之影響…....31
(三)、調查項目與記錄……………....….................32
(四)、生物統計分析…………………………………………….33
試驗結果………………………………………………….………..34
1. 供試藥材萃取物對爪哇根瘤線蟲二齡幼蟲的致死率及卵粒孵化
率之影響.………..…………………….…………….…...…...34
(1). 供試藥材20%水煮液對根瘤線蟲二齡幼蟲之致死率……….34
(2). 對葉百部新鮮塊根與乾燥塊根水煮液對根瘤線蟲二齡幼蟲致死率之影響..……………………………....….....…………....34
(3). 百部不同部位水煮液對根瘤線蟲二齡幼蟲致死率及卵粒孵化率之影響……………………..…………….…….......……...…35
(4). 百部之不同溶劑萃取物對根瘤線蟲二齡幼蟲致死率及卵粒孵化率之影響…………………………………………….............35
(5). 百部萃取物的pH值對根瘤線蟲二齡幼蟲致死率之影響....37
(6). 百部不同溶劑萃取物的貯放時間對根瘤線蟲二齡幼蟲致死率之影響……………………………………………………...........37
2. 百部不同溶劑之萃取物對爪哇根瘤線蟲防治效果與番茄TK70生育之影響…………………………………………………...…………38
(1). 百部不同溶劑之萃取物以不同施用量及不同施用時間對番茄根瘤線蟲之防治效果……………………………….........…..…38
(2). 百部不同溶劑之萃取物以不同施用量在不同週齡的番茄移植苗上施用對根瘤線蟲之防治效果………………….........……..40
(3). 百部不同溶劑之萃取物的施用次數對持效性之影響.…...42
結 論………………………………………………………….…..43
討 論……………………………………………………….……..44
參考文獻……………………………………………………….…..47
圖表………………………………………………………………...57
附件…………………………………………………………….…..98
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