一株耐盐溶磷真菌的筛选、鉴定及其生物肥料的应用效果  

Isolation and functional evaluation of phosphate-solubilizing fungi with salt-tolerant characteristics

作　　者：江红梅[1] 殷中伟[1] 史发超 刘彩月 程明芳[1] 范丙全[1] JIANG Hong-mei;YIN Zhong-wei;SHI Fa-chao;LIU Cai-yue;CHENG Ming-fang;FAN Bing-quan(Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences, Beijing 100081, China)

机构地区：[1]中国农业科学院农业资源与农业区划研究所,北京100081

出　　处：《植物营养与肥料学报》2018年第3期728-742,共15页Journal of Plant Nutrition and Fertilizers

基　　金：国家高技术研究发展计划(863计划)项目(2013AA102801-7);农业部"948"重点项目(2011-G25;2016-21X)资助

摘　　要：【目的】从内蒙古种植向日葵的盐碱地中筛选高效溶磷真菌,为农业生产中增产节肥,开发耐盐、溶磷微生物肥料提供菌种资源。【方法】利用形态特征和ITS r DNA序列鉴定菌株;LC-MS技术测定菌株M2在液体培养基中分泌有机酸和植物激素含量,明确菌株M2的溶磷和促生机理。采用液体摇床培养试验测定了鉴定菌株的溶磷能力。试验处理包括:在磷酸三钙、磷酸铝和5个磷矿的磷矿粉制备的100 m L难溶磷磷源(含5g/L难溶磷)中,接入1 m L灭菌培养液对照,和分别接种1 m L斜卧青霉菌P83和草酸青霉菌M2共15个处理。置于28℃、160 r/min摇床培养,分别于3、6和9 d,取菌液5 m L,在12000 r/min、4℃离心5 min,取上清液测定有效磷含量。采用含Na Cl的固体培养基测定菌株的耐盐性。Na Cl含量分别为0%、5%、7.5%、10%和12.5%的PDA平板中接入溶磷菌,置于28℃恒温培养箱中5 d,观察并记录菌丝的生长状况。采用盆栽试验方法检验了菌株的溶磷能力。以玉米种子(郑单958)为供试作物,以水稻土、黏性潮土、盐潮土和石灰性潮土为供试土壤,以Ca_3(PO_4)_2、AlPO_4和昆阳磷矿粉(RP)为供试磷源(磷源用量为1.0 g/kg土壤)。设置只加入灭菌草炭和Pikovskaya培养液对照,分别接种溶磷菌P83、M2,共计38个处理,144盆。玉米播种40天后收获,测定植株鲜重、干重和玉米根际土壤有效磷含量。田间试验以花生为供试作物,设置只加灭菌草炭和Pikovskaya培养液对照和分别接种ATCC20851、P83、M2溶磷菌剂三个处理。花生生长155 d后收获,称量花生植株鲜重和干重、花生果实鲜重和干重,同时采集花生根部土壤测定有效磷含量。【结果】溶磷菌株M2鉴定为草酸青霉(Penicillium oxalicum)。液体培养基摇床培养6 d后,接种菌株M2,以Ca_3(PO_4)_2为磷源的上清液中有效磷含量达972 mg/L,Ca_3(PO_4)_2溶解率为59.2%;以AlPO_4为磷源的有效磷含量达988 mg/L,溶解率�【Objectives】To supply effective microbial strains for biofertilizer production, we isolated phosphate-solubilizing fungi from the saline-alkaline soils planting sunflower in Inner Mongolia.【Methods】Petri dishes method was used to isolate phosphate-solubilizing fungus and the isolated fungus was identified by ITS r DNA sequence homology analysis. Broth medium culture was used to measure the dissolving capacity of insoluble phosphate of the isolated M2. Insoluble phosphates： Ca3（PO4）2, AlPO4, and five phosphorous rock powders as sole P source, respectively, were used to make PVK broth medium containing 5 g/L of phosphate separately, and 1 m L of sterilized cultural media, strain P83 and strain M2 were added into 100 m L of the broth medium and incubated at 28℃ for 3, 6 and 9 days at 160 r/min. Cultures were centrifuged at 12000 r/min for 5 min at 4℃ and the supernatant immediately analyzed for available phosphorus. To determinate the salt tolerance of isolate M2 and strain P83 in petridishes, the PDA solid medium containing 0%, 5%, 7.5%, 10% and 12.5% of Na Cl were employed and the growth diameters of P-solubilizing isolates P83 and M2 were recorded, respectively,after 5 days of incubation. A pot experiment was carried out to investigate solubilizing phosphate effect of strain M2. Maize seeds（Zea mays L. ‘Zhendan 958＇） were planted in the four types of soils（paddy soil, viscous fluvoaquic soil, salinized fluvo-aquic soil and calcareous fluvo-aquic soil） in the presence of Ca3（PO4）2, AlPO4 and Kunyang RP. The fresh and dry biomass and soil available phosphorus in treatments of P83 and M2 inoculations were measured at 40 days after sowing. A field trial was conducted to investigate the plant growth promotion effect of the produced bio-fertilizer with M2 strain. Peanut seeds were coated onto the mixture of fungal spores suspension（strain ATCC20851, P83 and M2） with sterilized peat. The yields of peanut straw and seeds were recorded and the soil available phosphorus was analyzed af

关 键 词：草酸青霉菌 溶磷作用 耐盐 促生效果 有机酸 植物激素 

分 类 号：S144[农业科学—肥料学]