臺灣博碩士論文加值系統

本研究旨在應用濾膜孢子發芽法探討影響叢枝菌根菌孢子發芽之因子，以及絲瓜接種叢枝菌根菌對根瘤線蟲之耐病性。孢子發芽試驗供試叢枝菌根菌有Acaulospora morrowiae (Am)、A. scrobiculata (As)、Entrophospora kentinensis (Ek)、Glomus aggregatum (Ga)、G. etunicatum (Ge)、G. mosseae (Gm)及Scutellospora spp. (Scu. spp.)等7菌種。4 ℃與室溫儲存對叢枝菌根菌Ge、Gm、Ga及As之孢子活力與發芽，在64週內並沒有明顯差異，除了Ga外，其餘三個菌種之孢子發芽率均隨著儲存時間增加而降低，As應用濾膜孢子發芽法對其發芽並不理想。比較一般河砂、蛭石、根基旺、滿地王及泥炭苔等介質對儲存2.5與1年之Ge、Gm、As及Ek孢子發芽影響，結果顯示不論儲存時間，Ge、Gm之孢子在蛭石之發芽率最高，As與Ek發芽情形則仍不理想，但是比較當年度生產並儲存三個月之Am、As、Ek、Ga、Ge及Gm，除了As、Gm外，其餘菌種孢子在蛭石之發芽率較高，在蛭石中可以發芽之菌種，發芽管之最大長度與分枝數目也較高。河砂顆粒大小與包埋深度對Ge菌種孢子發芽也有影響，包埋深度在1公分，以40孔目河砂之發芽率達28.3 %最高，其次為60孔目河砂達26.7 %，包埋深度在6公分，則以20 孔目河砂之發芽率達41.7 %為最高，其餘處理發芽率在20-21.7 %之間；Gm之孢子包埋於1公分深，發芽管長度會隨著河砂顆粒越細而減少，但分枝數目則增加，不論河砂顆粒大小，孢子以包埋於1公分深較包埋於6公分深有較高的發芽率；Am、As、Ek、Ga、Ge、Gm及Scu. Spp.等菌種之孢子，分別包埋於40孔目河砂、泥炭苔、3號根基旺及2號蛭石中，其中Am、Ek、Ga及Ge在40孔目河砂之發芽率最高，Gm則以在蛭石中發芽率最高，Scu. Spp.則以在3號根基旺有最高之發芽率，As在四種介質均未發芽。一般河砂、20、40、60孔目及2號蛭石分別作為繁殖Ge之介質，接種14週後百喜草菌根形成率，以一般河砂處理最高，其次為蛭石，但是每百毫升介質孢子數以蛭石最高達21,000個。Ek孢子包埋於一般河砂後，分別加入消毒之2號蛭石、3號根基旺、3號滿地王及泥炭苔過濾液，其中以添加3號根基旺與2號蛭石之過濾液，孢子發芽率分別為58與50.2 %，較對照組之46.1 %為高，加入滿地王與泥炭苔過濾液，發芽率則較對照組低，分別為21.2與30.5 %，但是發芽管分枝數目則以對照組最高達4.3枝，其次為添加蛭石過濾液之處理達3.9枝；河砂中硫銨添加量超過0.467 g/kg 、尿素2.18 g/kg 或磷礦石粉0.5 g/kg時明顯降低Gm孢子之發芽率，其中以增加硫銨用量抑制孢子發芽最明顯，過磷酸鈣用量在0.25 g/kg與1 g/kg，發芽率分別為16.7與18.3 %，與對照組之18.3 %沒有明顯差異，在Gm玉米盆栽試驗中，每公斤河砂分別添加1.09 g、1.63 g、2.17 g尿素與2.38 g、3.57 g、4.76 g硫銨，與添加1.5 g 3號奧妙肥及完全未添加肥料之處理進行產孢比較，接種後14週，以添加3.57 g與4.76 g硫銨之處理孢子數最高；同樣Gm玉米盆栽試驗，每公斤河砂分別添加2、4、6 g之過磷酸鈣與磷礦石粉，與添加1.5 g 3號奧妙肥及完全未添加肥料之處理進行產孢比較，其中以6 g磷礦石粉處理孢子量最高；3.57 g/kg硫銨、2.17 g/kg尿素、6 g/kg磷礦石粉單獨或混合施用，對盆栽玉米Gm產孢以單獨施用硫銨處理最高，其次為硫銨加磷礦石粉之處理，單獨施用尿素或尿素加磷礦石粉之處理則未有孢子產生；在盆栽百喜草試驗，以施用1.5 g/kg 3號奧妙肥為對照組，其孢子量最高每百公克達3823.3個，其次為單獨施用硫銨處理之2211.1個，單獨施用磷礦石粉之處理產孢最少僅276.7個。
叢枝菌根菌Am、As、Ek、Ga、Ge及Gm分別於絲瓜催芽後種植於穴盤並同時接種，2週及3週後，各菌種對絲瓜生長皆有促進效果；接種後6週，株高、地上部乾重、根系鮮重、根系乾重與感染率，以接種As之處理較佳；線蟲最初感染源密度試驗為接種200、400、600、800及1000隻二齡幼蟲，接種6週後結果顯示，各處理間根群生長有明顯差異，接種200隻即有根瘤形成，400隻則使根系根瘤數達30個以上，根鮮重為36.2 g和對照組26.2 g有顯著差異，故以400隻二齡幼蟲作為接種量；叢枝菌根菌與根瘤線蟲混合接種試驗，單獨接種線蟲者根系生長較少且根瘤數達50個以上，混合接種之植株根系皆較對照組與單獨接種線蟲者茂盛，罹根瘤數皆為30個以下，顯示對根瘤線蟲感染有減少之效果，接種Am、As、Ge及Gm菌種之植株，土壤中所含二齡幼蟲數量也相對減少；As與根瘤線蟲交互接種試驗中，亦以接種As者在株高及鮮重分別為201.5 cm及58.7 g最高，混合接種As及線蟲者之195.2 cm、54.1 g次之，根系生長以接種線蟲的根系鮮重8.28 g及乾重1.54 g最重，根瘤數均為50個以上，根瘤指數達4，而混合接種處理之根系乾重、根系鮮重分別為7.41 g和1.32 g，根瘤數30個以下，根瘤指數介於2－3之間，皆與對照組之6.76 g及1.19 g有顯著差異，每百公克乾土中南方根瘤線蟲二齡幼蟲數量，單獨接種線蟲者為16隻，較混合接種之9隻為高；於分根法（Split-root）試驗中得知，接種叢枝菌根菌促進絲瓜根系生長為系統性作用，兩邊分別接種As及線蟲者，根瘤數少於單邊接種線蟲者，且土壤中二齡幼蟲量亦有減少的現象。接種叢枝菌根菌使絲瓜根系中南方根瘤線蟲量減少之原因推測除了可能為侵染點之佔據，亦有可能為植株生理生化上的改變，因而使線蟲侵入減少，根瘤數亦減少，進而降低其土壤中之族群數量。

This study focused on the influence of factors on spores germination of arbuscular mycorrhizal fungi (AMF) by membrane filter technique, and the application of AMF for enhancing the tolerance of sponge gourd to root-knot nematode. There were 7 species of AM fungi to be twsted; that was Acaulospora morrowiae (Am), A. scrobiculata (As), Entrophospora kentinensis (Ek), Glomus aggregatum (Ga), G. etunicatum (Ge), G. mosseae (Gm) and Scutellospora spp. (Scu. spp.). Spore viability and germination of Ge, Gm, Ga, and As were not significantly different in 64 weeks stored in 4 ℃and room temperature after gathering. Percentage of spore germination rate decreased following the storage time increased in 4 species except Ga, and membrane filter technique was not adequate for germination of As. The effected of sand, vermiculite #2, king gardening #3 and flora pleur #3 on spore germination of As, Ek, Ge and Gm stored 1- and 2.5- year were compared, results indicated germination rate of Ge, Gm was the highest in vermiculite independented on storage time, and germination rate was still not well for As and Ek, but spore germination of Am, Ek, Ga and Ge stored for 3 months were higher in vermiculite than in other media except As and Gm, the length and branch of germination tube were higher in vermiculite than in other media. Particle size of sand and embedded depth affected the spore germination of Ge; embedded in 1 cm depth, the germination rate was 28.3 % in 40 mesh sand, next was 26.7 % in 60 mesh sand; embedded in 6 cm depth, germination rate was the highest, up to 41.7 % in 20 mesh sand, and 20-21.7 % in the other 3 treatments; for spores of Gm embedded in 1 cm depth, the length of germination tube decreased but branch increased following the particle size of sand decreased, germination rate of spores embedded in 1 cm depth was higher than in 6 cm depth independented of the size of sand. Germination rate of 7 species of AM fungi embedded in 4 media were compared, resulted indicated germination rate was higher for Am, Ek, Ga and Ge in 40 mesh sand, for Gm in vermiculite, for Scu. spp. in king gardening #3, and there was no germination for As in 4 media. Sand, 20 mesh, 40 mesh, 60 mesh sand, and vermiculite #2 were used for the propagation of Ge, colonization was the highest in sand at 14 weeks after inoculation, next was vermiculite, but spores per 100 ml medium was the highest and up to 21,000 in vermiculite. Filtration of vermiculite #2, KG #3, FP #3 and peat moss were added in sand where spores of Ek embedded, germination rate was 58 and 50.2 % in sand added with filtration of KG #3 and vermiculite #2, respectively, check was 46.1 %, and 21.2, 30.5 % for FP #3 and Peat moss, but the branch of germination tube was up to 4.3 for check, next was 3.9 for vermiculite filtration. Germination rate of Gm obviously decreased in sand when the amount was over 0.467 g, 2.18 g and 0.5 g/kg for ammonia sulfate, urea and rock phosphate, respectively; inhibition of germination was more obviously followed by the amount of ammonia sulfate increased, there was no significantly different in germination of Gm between check and those amended with 0.25, 1 g/kg of calcium superphosphate. For Gm propagation in corn pot culture, nitrogen amendment such as ammonia sulfate could increased the sporulation of Gm, but phosphate amendment such as rock phosphate or calcium superphosphate had no obvious increased, nitrogen and phosphate amended single or mix had different effected for Gm propagation in pot culture with corn or grass, ammonia sulfate was better for sporulation than other treatments in corn pot culture, but osmocote #3 was better in bahia-grass pot culture.
Growth promotion of sponge gourd inoculated with Am, As, Ek, Ga, Ge and Gm were compared, and growth of seedlings were promoted at 2 and 3 weeks after inoculation. Length and dry weight of shoot, fresh and dry weight of roots and colonization of seedlings inoculated with As were better than other treatments at 6 weeks after inoculation. The minimal inoculum density of root-knot nematode inducing disease symptom were tested, inoculum density were 0, 200, 400, 600, 800 and 1000 of second stage larvae of root-knot nematode in each pot. Results showed that the growth of root was significantly different in treatments at 6 weeks after inoculation. Root knots were induced when seedlings inoculated with 200 larvae per seedling, whereas there were more than 30 root knots in treatment of 400 larvae, fresh weight of root was 36.2 g and significantly different to control (CK) 26.2 g. The growth and disease incidenced of seedlings pre-inoculated with AMF for 2 weeks and then inoculated with 400 larvae of M. incognita were tested. There were over 50 root knots in seedlings inoculated with nematodes, and root growth of those inoculated with nematodes was less than control (CK). Root growth of seedlings co-inoculated with AMF and nematode was better than those of nematode alone, and root knots were less than 30. The results showed that sponge gourd pre-inoculated with AMF cound reduced the infection of M. incognita. The number of larvae per 100 g soil in treatments inoculated with Am, As, Ge and Gm also decreased. In co-inoculation tested of As and roo-knot nematode, length and fresh weight of shoots in treatment of As was 201.5 cm and 58.7 g, respectively, next was 195.2 cm and 54.1 g in treatment of As and Mi. Fresh and dry weight of root in treatment of Mi was 8.28 g and 1.54 g, respectively. It was more than 50 root knots in roots of Mi, and root knot index was up to 4, but fresh and dry weight in roots of As and Mi was 7.41 g and 1.32 g, respectively, the number of root knots was under 30 and root knot index was between 2-3. Fresh and dry weight of roots in Mi was significantly different with the control (CK) which was 6.76 g and 1.19 g. The number of 2nd larvae per 100 g soil in treatment of Mi was 16, and higher than the treatment of As and Mi which was 9. In split-root experiment, result showed that root growth promotion of sponge gourd inoculated with AM fungi was systemic. Root knots and 2nd larvae were less in seedlings which separately inoculated with As and root knot nematode in both sides than those inoculated with nematode in one side. Results indicated that sponge gourd inoculated with AM fungi might reduced the number of M. incognita due to occupation of infecting site, and physiological biochemistry of plants might change as well, therefore, the number of root knots and the population of 2nd larva in soil decreased.

中文摘要．．．．．．．．．．．．．．．．．．．．．．．．．．． I
英文摘要．．．．．．．．．．．．．．．．．．．．．．．．．．． IV
目次．．．．．．．．．．．．．．．．．．．．．．．．．．． VII
圖次．．．．．．．．．．．．．．．．．．．．．．．．．．． X
表次．．．．．．．．．．．．．．．．．．．．．．．．．．． XII
前言．．．．．．．．．．．．．．．．．．．．．．．．．．． 1
前人研究．．．．．．．．．．．．．．．．．．．．．．．．．．． 3
一、絲瓜簡介．．．．．．．．．．．．．．．．．．．．．．．．．．． 3
(一)生長特性．．．．．．．．．．．．．．．．．．．．．．．．．．． 3
(二)產業概況．．．．．．．．．．．．．．．．．．．．．．．．．．． 3
(三)功用．．．．．．．．．．．．．．．．．．．．．．．．．．． 3
(四)主要病害．．．．．．．．．．．．．．．．．．．．．．．．．． 4
二、根瘤線蟲簡介．．．．．．．．．．．．．．．．．．．．．．．．．．． 4
(一)分類地位．．．．．．．．．．．．．．．．．．．．．．．．．．． 4
(二)分佈及寄主範圍．．．．．．．．．．．．．．．．．．．．．．．．．．． 4
(三)生活史．．．．．．．．．．．．．．．．．．．．．．．．．．． 5
(四)病徵．．．．．．．．．．．．．．．．．．．．．．．．．．． 6
(五)防治方式．．．．．．．．．．．．．．．．．．．．．．．．．．． 6
三、叢枝菌根菌簡介．．．．．．．．．．．．．．．．．．．．．．．．．．． 6
(一)分類地位．．．．．．．．．．．．．．．．．．．．．．．．．．． 7
(二)形態構造．．．．．．．．．．．．．．．．．．．．．．．．．．． 7
(三)菌根之形成．．．．．．．．．．．．．．．．．．．．．．．．．．． 8
(四)影響孢子發芽及菌根感染之因子．．．．．．．．．．．．．．．．．．．．．． 9
(五)菌根形成對宿主植物之效益．．．．．．．．．．．．．．．．．．．．．．．．．． 11
材料與方法．．．．．．．．．．．．．．．．．．．．．．．．．．． 15
一、影響叢枝菌根菌孢子發芽與活力之因子．．．．．．．．．．．．．．．．．． 15
(一)供試叢枝菌根菌菌株來源與繁殖．．．．．．．．．．．．．．．．．．．．．．． 15
(二)試驗介質處理．．．．．．．．．．．．．．．．．．．．．．．．．．． 15
(三)孢子處理．．．．．．．．．．．．．．．．．．．．．．．．．．． 15
(四)孢子發芽觀察與活力測試．．．．．．．．．．．．．．．．．．．．．．．．．．． 16
(五)菌根形成率調查．．．．．．．．．．．．．．．．．．．．．．．．．．． 17
(六)儲存溫度與時間對叢枝菌根菌孢子活力及發芽之影響．．．．． 17
(七)介質種類對叢枝菌根菌孢子發芽之影響．．．．．．．．．．．．．．．． 17
(八)不同介質對6種叢枝菌根菌孢子發芽之影響．．．．．．．．．．．．．． 18
(九)河砂顆粒大小與包埋深度對叢枝菌根菌Ge及Gm孢子發芽之影響．．．．．．．．．．．．．．．．．．．．．．．．．．． 18
(十)40 mesh河砂及3種栽培介質對7種叢枝菌根菌孢子發芽之影響．．．．．．．．．．．．．．．．．．．．．．．．．．． 19
(十一)河砂顆粒大小與蛭石對Ge在百喜草上繁殖之影響．．．．．． 19
(十二)不同介質濾出液對叢枝菌根菌Ek孢子發芽之影響．．．．．． 19
(十三)氮、磷肥添加量對叢枝菌根菌Gm孢子發芽率之影響．．．． 20
(十四)氮、磷肥添加量對叢枝菌根菌Gm繁殖之影響．．．．．．．．．．． 20
(十五)氮、磷肥單獨或混合施用對叢枝菌根菌Gm於玉米與百喜草上繁殖之影響．．．．．．．．．．．．．．．．．．．．．．．．．．． 21
二、叢枝菌根菌與南方根瘤線蟲於絲瓜之交互作用．．．．．．．．．．．． 21
(一)供試植株．．．．．．．．．．．．．．．．．．．．．．．．．．． 21
(二)供試叢枝菌根菌菌種．．．．．．．．．．．．．．．．．．．．．．．．．．． 22
(三)供試南方根瘤線蟲蟲源及其培養．．．．．．．．．．．．．．．．．．．．．．． 22
(四)根組織透化染色及叢枝菌根菌感染率之調查方法．．．．．．．．． 22
(五)不同叢枝菌根菌對絲瓜苗生長促進效果之比較．．．．．．．．．． 23
(六)南方根瘤線蟲最適接種量測定．．．．．．．．．．．．．．．．．．．．．．．．． 23
(七)不同叢枝菌根菌與南方根瘤線蟲單獨或混合接種對絲瓜生長之影響．．．．．．．．．．．．．．．．．．．．．．．．．．． 24
(八)叢枝菌根菌A. scrobiculata與南方根瘤線蟲單獨或混合接種對絲瓜生長之影響．．．．．．．．．．．．．．．．．．．．．．．．．．． 25
(九)應用分根法測定叢枝菌根菌A. scrobiculata與南方根瘤線蟲在絲瓜之相互作用．．．．．．．．．．．．．．．．．．．．．．．．．．． 25
結果．．．．．．．．．．．．．．．．．．．．．．．．．．． 26
一、影響叢枝菌根菌孢子發芽與感染之因子．．．．．．．．．．．．．．．．．． 26
(一)儲存溫度與時間對叢枝菌根菌孢子活力及發芽之影響．．．．． 26
(二)不同種叢枝菌根菌儲存時間對其孢子活力及發芽之影響．．．． 31
(三)介質種類對叢枝菌根菌孢子發芽率之影響．．．．．．．．．．．．．．． 31
(四)河砂顆粒大小及包埋深度對叢枝菌根菌Ge和Gm孢子發芽之影響．．．．．．．．．．．．．．．．．．．．．．．．．．． 35
(五)40 mesh河砂及3種栽培介質對7種叢枝菌根菌孢子發芽率之影響．．．．．．．．．．．．．．．．．．．．．．．．．．． 35
(六)叢枝菌根菌Ge於不同顆粒大小河砂與蛭石中之繁殖比較．．． 40
(七)不同介質濾出液對叢枝菌根菌Ek孢子發芽之影響．．．．．．．． 40
(八)氮、磷肥添加量對叢枝菌根菌Gm孢子發芽率之影響．．．．．．． 44
(九)氮、磷肥添加量對叢枝菌根菌Gm繁殖之影響．．．．．．．．．．．．． 44
(十)氮、磷肥單獨或混合施用對叢枝菌根菌Gm於玉米與百喜草上繁殖之影響．．．．．．．．．．．．．．．．．．．．．．．．．．． 51
二、叢枝菌根菌與南方根瘤線蟲於絲瓜之交互作用．．．．．．．．．．．． 54
(一)不同叢枝菌根菌對絲瓜苗之生長促進效益．．．．．．．．．．．．．．． 54
(二)南方根瘤線蟲之最適接種量．．．．．．．．．．．．．．．．．．．．．．．．．．． 60
(三)不同叢枝菌根菌與南方根瘤線蟲混合接種對絲瓜生長之影響．．．．．．．．．．．．．．．．．．．．．．．．．．． 65
(四)叢枝菌根菌A. scrobiculata與南方根瘤線蟲單獨或混合接種對絲瓜生長之影響．．．．．．．．．．．．．．．．．．．．．．．．．．． 72
(五)應用分根法測定叢枝菌根菌A. scrobiculata與南方根瘤線蟲在絲瓜之相互作用．．．．．．．．．．．．．．．．．．．．．．．．．．． 72
討論．．．．．．．．．．．．．．．．．．．．．．．．．．． 82
結論．．．．．．．．．．．．．．．．．．．．．．．．．．． 89
參考文獻．．．．．．．．．．．．．．．．．．．．．．．．．．． 91

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