根际促生菌株的筛选及其复合微生物肥料的研制与肥效研究

发布时间：2018-04-10 10:06

  本文选题：根际促生菌　切入点：IAA　出处：《南京农业大学》2015年硕士论文

【摘要】：由于长期不合理施用化肥,我国耕地土壤结构和土壤微生物群落结构遭到严重破坏,化肥的增产效果呈逐年降低趋势,农产品品质下降,化肥利用效率偏低进而造成环境污染。随着生产力和生活水平的提高,单施化肥已经不能满足人们对绿色农业的需求。然而传统的有机肥有效养分低,肥效慢,施用量大,造成大量资源损耗,经济效益低。由根际促生菌制成的生物菌肥效率低、应用规模小、无区域针对性。将生物菌肥与有机肥和化肥有机结合,综合三者优势开发新型肥料具有广阔的应用前景。本研究首先分离筛选出能产IAA和ACC脱氨酶的菌株。结果显示,NJAU-52、NJAU-FAD、NJAU-60、NJAU-69和NJAU-29五株菌产IAA的量较高,产量为7.80～14.90 mg·L~(-1)。菌株 NJAU-54、NJAU-82、NJAU-69、NJAU-53、NJAU-84、NJAU-72、NJAU-86能产ACC脱氨酶,其酶比活力为0.18～0.45 U·mL~(-1)·min~(-1)。综合以上实验结果筛选出 NJAU-FAD、NJAU-25、NJAU-29、NJAU-52、NJAU-53、NJAU-54、NJAU-57、NJAU-60、NJAU-69、NJAU-84 这 10 株菌进行灌菌盆栽试验。通过盆栽试验结果,复筛获得4株促生效果较佳的菌株,分别为NJAU-52、NJAU-57、NJAU-60和NJAU-69,经鉴定,分别为内生芽孢杆菌(Bacillus endophyticus)、弯曲芽孢杆菌(Bacillus flexus)、海水芽孢杆菌(Bacillus aquimaris)和巨大芽抱杆菌(Bacillus megaterium)。选择上述4株菌株进行第二季灌菌盆栽试验以验证促生功能的稳定性,最后获得促生效果最优的菌株NJAU-60。通过灌施菌株NJAU-60能使玉米植株株高增加20%、地上部鲜重增加32%、地上部干重增加20%;使黄瓜植株株高增加10%、地上部干重增重12%;使茄子植株株高增加52%、叶绿素值增加6.3%;使番茄植株株高增加16%、茎粗增加16%、叶绿素增加13%、地上部鲜重增加33%、地上部干重增加20%。利用菌株NJAU-60和本实验室保存的根际促生菌Bacillus amyloliquefaciens SQR-9分别研制成粉状和颗粒状复合微生物肥,并在存储期间对颗粒复合微生物肥中功能菌株涂布计数。含菌株NJAU-60复合微生物肥料在存储至180 d时原养分和7%养分的肥料中菌株NJAU-60的数量分别为19.5 × ~(-1) cfu·g~(-1)和7.3 × ~(-1) cfu·g~(-1),高于国家标准(≥2.0 × ~(-1) cfu·g~(-1)),10%养分的肥料中菌株NJAU-60的数量为2.1 × ~(-1) cfu·g~(-1),正好达到国家标准(≥2.0×~(-1)cfu·g~(-1)),15%养分和20%养分的肥料中菌株NJAU-60 的数量分别为 0.8×~(-1) cfu·g~(-1) 和 0.2 × ~(-1) cfu·g~(-1),低于国家标准(≥2.0 × ~(-1) cfu·g~(-1))。含菌株SQR-9复合微生物肥料保存至180 d时,10%养分和15%养分肥料中菌株SQR-9的数量分别为11.33 × 1 07 cfu·g~(-1),15养分为7.8 × 1 07 cfu·g~(-1),均达到国家标准(≥2.0×~(-1) cfu·g~(-1)。对含菌株SQR-9复合微生物肥料的粉状肥料和颗粒肥料进行养分释放试验和盆栽试验。养分释放试验结果表明粒状肥的养分释放慢于粉状肥,到第60 d时,10%养分粉状和颗粒复合微生物肥料重量分别减少了 22.7%和17.4%;15%养分粉状和颗粒复合微生物肥料重量分别减少了 32.3%和22%。在盆栽试验中,等养分的有机无机复混肥处理比化肥处理的植株长势更好,粉状复合微生物肥料处理比有机无机复混肥处理的植株长势更好,颗粒复合微生物肥料处理与粉状复合微生物肥料处理的植株长势一致。
[Abstract]:Due to the long-term unreasonable application of chemical fertilizer, the severe destruction of China's arable land soil structure and soil microbial community structure, chemical fertilizer yield decreased year by year, decline in the quality of agricultural products, fertilizer utilization efficiency is low and cause environmental pollution. With the development of productive forces and the improvement of living standards, chemical fertilizer has been unable to meet the demand for green agriculture however the traditional organic fertilizer nutrient fertilizer application rate is low, slow, resulting in a large number of resource depletion, low economic benefits. By promotingrhizobacteria made of bio fertilizer application in small scale, low efficiency, no region specific. Bio fertilizer and organic fertilizer and chemical fertilizer combined with organic, integrated the advantages of three development a new type of fertilizer has a broad application prospect. This study first isolated can produce IAA and ACC deaminase strains. The results showed that NJAU-52, NJAU-FAD, NJAU-60, NJAU-69 and NJAU-29 were five Bacteria that produce higher levels of IAA, the yield is 7.80 ~ 14.90 Mg - L~ (-1). NJAU-82, strain NJAU-54, NJAU-69, NJAU-53, NJAU-84, NJAU-72, NJAU-86 can produce ACC deaminase, the enzyme activity was 0.18 ~ 0.45 U - mL~ (-1) - min~ (-1). The above results were NJAU-FAD, NJAU-25, NJAU-29, NJAU-52, NJAU-53, NJAU-54, NJAU-57, NJAU-60, NJAU-69, NJAU-84 of the 10 strains of bacteria by pot experiment. Through pot experiment, 4 strains were screened for promoting better results, respectively NJAU-52, NJAU-57, NJAU-60 and NJAU-69, were identified, in Bacillus (Bacillus endophyticus), b.flexus (Bacillus flexus), seawater bacillus (Bacillus aquimaris) and Bacillus megaterium (Bacillus megaterium). The 4 strains of bacteria were second season pot experiment to verify the stability of the promoting function, finally get the optimal growth promoting effect The strain NJAU-60. by strain NJAU-60 can make the irrigation and fertilization of maize plant height increased 20%, the fresh weight of shoot dry weight increased by 32%, an increase of 20%; the plant height increased 10%, shoot dry weight of 12%; the eggplant plant height increased 52%, SPAD value increased by 6.3%; the tomato plant high increased 16%, stem diameter increased 16%, 13% increase in chlorophyll, shoot fresh weight, shoot dry weight increased by 33%, 20%. increased by NJAU-60 strains preserved in our laboratory and Rhizosphere growth promoting bacteria Bacillus amyloliquefaciens SQR-9 were developed into powder and granular composite microbial fertilizer, and during the storage of granular composite microbial fertilizer in function strains containing strain NJAU-60. Count coated composite microbial fertilizer in storage at 180 d and 7% primary nutrient fertilizer amount of strain NJAU-60 were 19.5 x ~ (-1) CFU - g~ (-1) and 7.3 ~ (-1) CFU * g~ (-1), higher than China 瀹舵爣鍑，

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