【作者】 杨小明；

【导师】 叶建仁；

【作者基本信息】 南京林业大学 ， 森林保护， 2004， 硕士

【摘要】 松树枯梢病(Sphaeropsis sapinea)这一世界性松树病害在我国南方地区引起大面积松林衰退或死亡，使大面积的国外松幼林遭受毁灭性损失。近十年来，如何控制松树枯梢病这一林木寄主主导性病害受到高度的重视。目前国内主要通过提高林木个体、群体抗逆性、调节林分和植株的微生态环境，改善林木根系、松树菌根以及根际微生物区系等方面来研究南方国外松人工林的生态控制技术。本文的立意就是通过引入松树菌根，提高寄主的生长势，以达到防治病害的目的。 本文通过对松树外生菌根真菌的采集、分离，培养筛选；菌剂制备和接种效应分析等，确立了彩色豆马勃、黄须腹菌对湿地松幼苗的共生关系，同时发现彩色豆马勃与湿地松在室内接种条件下形成内外生菌根 1、从云南的6个不同采集点采集到7个科，11个属，13个种的外生菌根真菌，通过菌肉组织分离法、菌索分离法、单孢分离法分离到彩色豆马勃(Pisolithus tinctorius (Pers．)Coker et couch)、须腹菌(Rhizipogen luteous)、黄豹斑鹅膏(Amanita pantherina)三种松树的外生菌根真菌。 2、室内实验对采集分离和收集到的分属于10个属的11种外生菌根真菌进行培养条件的摸索，并选取培养状况好的菌株进行接种试验，两株彩色豆马勃的最佳培养基分别为综合PDA和Bain培养基，并在27℃—30℃的培养条件下快速生长。选出两株彩色豆马勃、黄须腹菌、黄豹斑鹅膏这4株菌作为进一步试验菌株。 3、针对不同菌株摸索出的培养基配方及培养条件，并利用固体培养和液体培养制备出4株菌根真菌的菌剂接种体，固体培养的菌根菌剂在一个月后制备成功，但不同菌种之间有一定差异，以两株彩色豆马勃菌株制备的菌剂长势最好。 4、用菌根真菌接种湿地松无菌苗确定两者之间的共生关系。根据不同的菌根菌剂，不同的菌剂剂型和不同的菌剂施用量接种湿地松幼苗，分析各种不同试验设计下的菌根菌的感染率。 5、菌根化育苗及菌根菌与宿主植物共生关系的研究中，发现实生湿地松幼苗在接种外生菌根真菌1个半月左右后，在幼苗的根部形成不同程度的菌丝套结构，接种彩色豆马勃和黄须腹菌形成了二叉状分支的菌根。 6、接种所有供试菌剂而形成的菌根，制成石蜡切片后均观察到不同程度的哈缔氏网结构和菌丝套结构，其中尤其在接种彩色豆马勃2个月后形成的菌根中发现，菌丝体已经侵入到宿主植物根部的皮层细胞，这证明了彩色豆马勃不仅能在根表面形成菌丝套并侵入到根部细胞的细胞间隙形成哈缔氏网，而且能深入到皮层细胞内形成内外生菌根。更多还原

【Abstract】 Pine shoot blight and canker is the pine tree disease of world harms to cause big area pine wood decline or death in the southern region in our country, making the abroad pine of the big area the wood suffered from the destructive loss. In the last decade, how to control the influence of Pine shoot blight and canker in pine tree (Host leading diseases) to our country forestry was valued highly. The chief ecological approachs to control prevalence and occurrence are to enhance the whole anti- adversity ability and disease resistance by improveing rhizosphere micro-bial environment and micro- ecological environment.Through collecting , isolateing and cultivating the ectomycorrhizal fungi, this experiment produced the mycorrhiza agent and analysised the effect of inoculate Pinus elliottii .making sure the Pisolithus tinctor-ius(Pers)Coker et Couch, Rhizipogen luteous to the symbiosis relation of the Seedlings of Pinus elliottii. At the same time Discovers the ect-endomycorrhiza characteristic of Pisolithus tinctorius(Pers) Coker et Couch. The main results as follows:1, 13 species mycorrhiza fungi were collect from Yunnan . Pisolithus tinctorius(Pers)Coker et Couch, Rhizipogen.sp and Amanita pantherina(Dc: Fr)Secr were isolated form all the 13 species mycorrhiza fungi by using methods of isolating context of fungi; methods of isolating rhizomorph ; methods of isolating single spore.2, Culture conditional sieving among 11 strians mycorrhiza fungi ,4 strains mycorrhiza fungi were selected for the lately stage Inoculating experiments .3 , Four types mycorrhizas agent were produced by methods of solid culture and methods of liquid culture. Solid mycorrhizas agent was produced after 1.5 month, liquid mycorrhizas agent was produced after 10 day .all testing strain had different culture characteristics. Two strains Pisolithus tinctorius (Pers)Coker et Couch have good culture status .4, Making sure the symbiosis relation between the mycorrhiza fungi and host plant by using the mycorrhiza fungi inoculating the asepsis seedlings of Pinus elliottii.The. rate of inoculation was analysised under different mycorrhizas agent ;medicament form and agent usage.5,The symbiosis relation are ascertain by the mycorrhiza formed after inoculated for 1.5 month . Mycelium were found in the root of all the 4 type inoculatedseedlings.Two arrows limb were found on the mycorrhiza of P. elliottii which inoculated by Pisolithus tinctorius(Pers)Coker et Couch and Rhizipogen luteous.6, Analysis the micro structure of mycorrhizas by paraffin microtome method. Hypha can be found have infest into the exodermis of host plant after inoculated by Pisolithus tinctorius(Pers)Coker et Couch for 2 months.The paraffin section have proved the hypha of Pisolithus tinctorius (Pers)Coker et Couch not only formed the mycelium outside of the root and percolate into the interspace of root cell ,but also penetrating into the cell of exodermis .the structure of ectoendotrophic mycorrhiza are formed by P.tinctorius(Pers)Coker et Couch更多还原