• ISSN 1008-505X
  • CN 11-3996/S
冯程龙, 王晓婷, 康文晶, 孟晓慧, 张风革, 冉炜, 沈其荣. 利用小麦秸秆生产木霉分生孢子及其生物有机肥对黄瓜的促生效果[J]. 植物营养与肥料学报, 2017, 23(5): 1286-1295. DOI: 10.11674/zwyf.17098
引用本文: 冯程龙, 王晓婷, 康文晶, 孟晓慧, 张风革, 冉炜, 沈其荣. 利用小麦秸秆生产木霉分生孢子及其生物有机肥对黄瓜的促生效果[J]. 植物营养与肥料学报, 2017, 23(5): 1286-1295. DOI: 10.11674/zwyf.17098
FENG Cheng-long, WANG Xiao-ting, KANG Wen-jing, MENG Xiao-hui, ZHANG Feng-ge, RAN Wei, SHEN Qi-rong. Trichoderma conidia production using wheat straw and growth promoting effect of its bio-organic fertilizer on cucumber[J]. Journal of Plant Nutrition and Fertilizers, 2017, 23(5): 1286-1295. DOI: 10.11674/zwyf.17098
Citation: FENG Cheng-long, WANG Xiao-ting, KANG Wen-jing, MENG Xiao-hui, ZHANG Feng-ge, RAN Wei, SHEN Qi-rong. Trichoderma conidia production using wheat straw and growth promoting effect of its bio-organic fertilizer on cucumber[J]. Journal of Plant Nutrition and Fertilizers, 2017, 23(5): 1286-1295. DOI: 10.11674/zwyf.17098

利用小麦秸秆生产木霉分生孢子及其生物有机肥对黄瓜的促生效果

Trichoderma conidia production using wheat straw and growth promoting effect of its bio-organic fertilizer on cucumber

  • 摘要:
    目的木霉是重要的植物根际促生功能菌,然而木霉类生物肥料生产的应用基础研究很薄弱。本研究以小麦秸秆为发酵基质,探究哈茨木霉T-E5产分生孢子的固体发酵工艺条件,并验证发酵物作为生物有机肥的促生效果。
    方法在实验室条件下,以小麦秸秆为固体发酵基质,研究固体发酵的不同发酵方式,温度、酸浓度、酸种类、接种量、含水量、外加碳源和氮源等单因子条件下的产孢量;再利用正交试验设计,研究温度、酸浓度、含水量、接种量、外加氮源和碳源的发酵条件优化。利用两季温室盆栽试验测定包括施用化肥、有机肥、发酵物和不同肥料组合的5个不同施肥处理的黄瓜地上部生物量、株高和叶面积。采用稀释涂布法测定黄瓜盆栽根际土中总真菌数量和木霉菌数量。
    结果单因素条件优化:基质用稀硝酸 (0.1 mol/L) 调节酸度值后接种3%(v/w) 的1 × 107个/mL的T-E5孢子液,调节水分使初始含水量达到75%;在此条件下,添加尿素和玉米粉,在28℃下发酵培养8天,最大产孢量可达到8.87 × 109个/g。通过正交优化,每10 g小麦秸秆,加入调节酸度值的0.05 mol/L硝酸18.3 mL,含水量70%(湿基),接种量9%,添加尿素的量为1%,30℃浅盘培养8天,产孢量达到1.1 × 1010个/g。第1季盆栽试验结果表明,与单施化肥处理 (CCK) 相比,发酵产物与化肥配施 (CT) 使黄瓜的地上部分生物量、株高和叶面积分别提高14.97%、16.75%和14.64%;与单施有机肥处理 (OCK) 相比,发酵产物与有机肥配施 (OT) 使黄瓜的地上部分生物量、株高和叶面积分别提高10.62%、26.06%和9.53%。第2季盆栽也有类似的促生效果,与CCK相比,CT使上述指标分别提高12.83%、11.49%和26.93%;与OCK相比,OT使上述指标分别提高5.96%、34.29%和18.01%。两季盆栽添加木霉发酵物的处理中根际土总真菌数和木霉菌数量相比CCK、OCK和不施肥照 (CK) 均增加1个数量级以上。
    结论以小麦秸秆为主要原料,采用固体发酵工艺可生产高分生孢子含量的哈茨木霉菌剂及其生物有机肥,表明利用农业废弃物生产高附加值的木霉菌剂及其生物有机肥具有可行性。但木霉孢子数量主要受发酵条件影响,今后应在中试和工厂化生产水平进一步研究木霉孢子固体发酵的工艺参数。

     

    Abstract:
    ObjectivesTrichoderma species are important plant growth promotion fungi in rhizosphere of plants, whereas it is insufficient for using them to produce bio-fertilizers. This study tried to use wheat straw powder as substrate for conidia production of T. harzianum T-E5 in solid state fermentation (SSF), aimed to optimize process conditions of the fermentation as well as to check the plant growth promoting effect of the ferment produce as a bio-organic fertilizer on cucumber in greenhouse.
    MethodsIn laboratory conditions, wheat straw was used as solid fermentation substrate, and numbers of conidia spores were compared under a series of single factors, including different fermenting containers, temperatures, acid concentration, acid species, amounts of inoculum, moisture contents, and forms of exogenous carbon and nitrogen resources. An orthogonal experiment was then designed to optimize fermentation conditions of temperature, acid concentration, water content, inoculation amount, and external nitrogen and carbon sources. In addition, two seasons of greenhouse pot experiments were conducted with five fertilization treatments, including chemical fertilizer, organic fertilizer, ferment product and its combinations with the chemical fertilizer or the organic fertilizer. The above ground biomass, plant heights and leaf areas of cucumber were measured, The number of total fungi and Trichoderma species in the rhizosphere soil of cucumber were counted using the diluted spread plate.
    ResultsThe solid state fermentation of T-E5 in tray with wheat straw powder was suitable for production of conidial spores. The wheat straw powder was prepared by inoculation with 3% (v/w) of 1 × 107 spores/mL T-E5 spore suspension prior to acidification with nitric acid solution (0.1 mol/L) and then water was added to reach an initial moisture content of 75%. Under those conditions, fermentation with addition of exogenous urea and corn powder at 28℃ for 8 days yielded the maximum number of T-E5 conidial spores of 8.87 × 109 spores/g. By the orthogonal optimization, when 18.3 mL 0.05 mol/L of nitric acid, 9% (v/w) inoculation amount, and 1% (w/w) of urea were added to every 10 g of wheat straw with 70% (wet basis) of water content, at 30℃ shallow dish and fermented for 8 days, the amount of Trichoderma conidia reached to 1.1 × 1010 spores/g. The results of the first season pot experiment showed that the treatment of ferment product with chemical fertilizers (CT) increased cucumber above ground biomass, plant height and leave area by 14.97%, 16.75% and 14.64%, respectively, compared with the treatment of chemical fertilizers (CCK). Compared with the treatment of the organic fertilizer (OCK), the treatment of ferment product with the organic fertilizer (OT) increased the above ground biomass, plant height and leave area by 10.62%, 26.06% and 9.53%, respectively. The results of the second season pot experiment showed similar promoting effects, i.e. compared with CCK, CT increased the above items by 12.83%, 11.49% and 26.93% respectively, and compared with OCK, OT increased above items by 5.96%, 34.29% and 18.01%, respectively. Compared with CCK, OCK and CK, the CT and OT treatments increased the numbers of total fungi and Trichoderma species in the rhizosphere soil of cucumber by more than one order of magnitude.
    ConclusionsThe bio-agent and bio-organic fertilizer with high content of T. harzianum conidial spores can be produced using wheat straw as main substrates in SSF process. This suggests it is feasible that using agricultural wastes with SSF process to produce high value added Trichoderma bio-agent and its bio-organic fertilizers. But the amount of spores is largely dependent on the fermentation conditions, and the technological parameters of SSF for producing Trichoderma spores in the pilot and factory levels need to be further studied.

     

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