盤固拉草堆肥添加康氏木黴菌防治甘藍幼苗立枯病之探討

Antagonistic activity of selected strains of Trichoderma koningii amended in compost of pangolagrass against Rhizoctonia solani, pathogen of cabbage seedling damping-off

李柏賢(Bo-Shian Li)

指導教授 ： 梁文進

國立屏東科技大學/農學院/植物醫學系所/碩士(2009年)

https://doi.org/10.6346/NPUST.2009.00317

摘要

本研究探討以木黴菌或添加至盤固拉草堆肥中，試驗其防治甘藍苗立枯病之效果。由屏東地區土壤、植物枯枝上分離29株木黴菌菌株，對甘藍立枯病菌(Rhizoctonia solani AG-4)，以玻璃紙抗生法及對峙培養篩得TML、TSR、TCL等木黴菌菌株。該三菌株亦可抑制蓮霧黑腐病菌(Lasiodiplodia theobomae)、果腐病菌(Pestalotiopsis euginae)、黃腐病菌(Cylindrocladium sp.)、印度棗炭疽病菌(Colletotrichum gloeosporioides)、茄子疫病菌(Phytophthora capsici)、蔥白絹病菌(Sclerotium rolfsii)菌絲生長。利用顯微鏡觀察雙層載玻片對峙培養，發現木黴菌TML、TSR、TCL菌株可造成R. solani病原菌菌絲斷裂及穿生現象。利用核醣體DNA比對鑑定TML、TSR、TCL菌株皆為Hypocera koningii(anamorph: T. koningii)。於盤固拉草堆肥化過程，經測其溫度、pH值，電導度EC值，微生物族群變動，微生物活性及對種子發芽率，獲知堆積發酵35天候，就可獲得成熟的盤固拉草堆肥。盤固拉草堆肥萃取液培養基中添加米糠或麥麩，可以促進木黴菌菌絲生長;添加米糠、全麥粉、黃豆粉可促進產孢達1.44×108 spores/g，其產生孢子發芽率達92%以上。將8種營養物添加於盤固拉草堆肥中測試其菌絲生長情形，發現TML、TCL、TSR於添加麥麩、米糠、小麥粉時，木黴菌菌絲生長最佳，但於添加甲殼素堆肥中生長之菌絲較其他添加物者差。添加9%培養木黴菌TML之穀粒於病土中，可完全抑制甘藍幼苗立枯病之發生。而使用5%培養木黴菌TSR之穀粒，就可完全抑制立枯病之發生。添加10%培養木黴菌TML及TSR菌株之盤固拉草堆肥於病土中，可分別獲得甘藍苗立枯病防治率為21及33%，而添加20%時，則防治率分別為73及82%。

關鍵字

康氏木黴菌 ； 盤固拉草堆肥 ； 甘藍苗立枯病 ； 微生物防治

並列摘要

The purpose of this research was focused on ultilization of Trichoderma antagonists alone or amended in pangolagrass compost to test the control efficiency of cabbage seedling damping-off. A total of 29 isolates of Trichoderma spp. were isolated from soils and dead materials of plant at different areas in Pingtung. With cellophane antagonism test and dual culture, the antagonistic activity of these Trichoderma isolates was tested, and 3 isolates of Trichoderma spp. were selected based on the inhibitory effect on mycelial growth of Rhizoctonia solani AG-4, pathogen of cabbage seedling damping-off. These three antagonistic Trichoderma isolates, designated TML, TSR and TCL, could also inhibit mycelial growth of Lasiodiplodia theobomae, Pestalotiopsis euginae. Cylindrocladium sp., Colletotrichum gloeosporioides, Phytophthora capsici and Sclerotium rolfsii. Dual culture on slide between Trichoderma sp. and R. solani AG-4 showed that TML, TSR and TCL isolates could degrade the mycelium of R. solani; in addition, the hypha of Trichoderma isolate TCL could penetrate into hypha of R. solani. All of three isolates were identified as species of Hypocera koningii (anamorph: Trichoderma koningii) by comparation of the sequence of ribosomal DNA. In composting process of pangolagrass, through monitory changes of the temperature, pH value, electronic conductivity, population dynamic of microognisms, microbial activity and seed germination test in extract of compost, a mature compost could be made after 35 days fermentation. On the agar medium plates made of extract of pangolagrass compost amended with various nutrients, the mycelial growth of the T. koningii isolates was increased when rice bran or wheat bran was amended; and the sporulation was also increased when rice bran, whole wheat flour, or soybean flour was amended. The spores produced from culture of compost extract had spore germination rate of 92% or more. Isolates TML, TCL and TSR could grow well on pangolagrass compost amended with wheat bran, rice bran or wheat flour, but not with chitin amendment. When 9% wheat grain which had been grown T. koningii TML isolate was amended to diseased soil that was infested with 5% R. solani culture materials, T. koningii TML could control 100% damping-off, while 5% T. koningii TSR could control damping-off completely. When R. solani infested soil was added with 10% pangolagrass compost which had been grown T. koningii TML or TSR isolate, the control efficiency of seedling damping-off was at 21 or 33% respectively, whereas with 20% compost amendment, 73 and 82% of control efficiency, respectively.

並列關鍵字

Trichoderma koningii ； pangolagrass compost ； Rhizoctonia solani ； microbial control

參考文獻

林景和。1993。利用廢棄菇類栽培介質製作堆肥之研究。臺中區農業改良場研究彙報39：17-27。

林財得、謝清祿、謝欽城。2004。不同前處理對稻桿吸水性之影響及以稻桿與禽畜糞製作堆肥可行性探討。農業機械學刊13:1-12。

Huang, J. W., and Huang, H. C. 2000. A formulated container medium suppressive to Rhizoctonia damping-off of cabbage. Bot. Bull. Acad. Sin. 41:49-56.

陳純葳。2006。土壤添加蝦蟹殼粉防治番茄萎凋病之效果。國立中興大學植物病理學系碩士論文。48頁。

楊正偉。2006。苦茶粕防治甘藍幼苗立枯病之效果與其有效成分之鑑定。國立中興大學植物病理學系碩士論文。49頁。