The contribution of endogenous cellulase to the cellulose digestion in the gut of earthworm (Pheretima hilgendorfi: Megascolecidae)

Cellulase activity has been detected in the digestive tract of earthworms. However, it has not been well clarified whether the origin of those cellulases are the earthworm themselves or the symbionts. In our study, zymogram analysis suggests that one cellulase (endo-β-1,4-glucanase, EC3.2.1.4) mainly works to digest cellulose in Pheretima hilgendorfi. To identify the cellulase in P. hilgendorfi, we carried out cDNA cloning of the cellulase gene from the digestive tract. A novel cellulase gene was identified from the gut of earthworm. The cDNA encoding cellulase of P. hilgendorfi (phhEG) is 1606 bp with an open reading frame encoding a protein of 449 amino acid residues. The deduced amino acid sequence of P. hilgendorfi cellulase showed higher homology to invertebrate cellulases than bacterium cellulases belonging to the glycosyl hydrolase family (GHF) 9. The phhEG gene was detected in intestinal epithelium cell of midforegut using Northern blot and in situ hybridization. Similarly, specific cellulase activity against carboxymethyl cellulose (CMC) was significantly higher in midforegut tissue. Recombinant phhEG produced by wheat germ cell-free protein synthesis system had a cellulase activity which degrade CMC. In zymogram analysis, the molecular size of cellulase was detected as a single band of 51 kDa from the whole gut contents extracts of P. hilgendorfi, and was very similar to the predicted molecular size of the mature phhEG protein. These results strongly suggested that the earthworm has the capacity to produce the endogenous and functional cellulase around the midforegut, and use this cellulase for their cellulose digestion with the support of intestinal caecum.     

蚕豆根瘤菌的分离、筛选及其肥效研究

从甘肃省不同蚕豆种植区采集蚕豆根瘤,采用选择性培养基,分离获得到11株蚕豆根瘤菌,经过蛭石粉和土壤栽培筛选试验,筛选出7株(G1、G2、G6、G7、G8、G10、G11)根瘤菌,它们与临蚕2号蚕豆有效共生匹配效果好,共生固氮力强,具有一定的生产应用价值。     

丛枝菌根和根瘤菌双接种对矿区土地复垦的生态效应

通过盆栽试验研究了山西省襄垣县矿区复垦土壤中接种丛枝菌根真菌和根瘤菌对三叶草的生长状况及土壤养分的影响.结果表明,双接种丛枝菌根和根瘤菌能显著提高菌根侵染率和土壤中孢子密度,促进三叶草干物质的积累和对氮、磷元素的吸收,促进三叶草对土壤养分的活化,提高土壤养分利用率,加速矿区生态恢复.     

Analysis of Simple Sequence Repeats in Genomes of Rhizobia

Simple sequence repeats （SSRs） or microsatellites, as genetic markers, are ubiquitous in genomes of various organisms. The analysis of SSR in rhizobia genome provides useful information for a variety of applications in population genetics of rhizobia. We analyzed the occurrences, relative abundance, and relative density of SSRs, the most common in Bradyrhizobium japonicum, Mesorhizobium loti, and Sinorhizobium meliloti genomes se- quenced in the microorganisms tandem repeats database, and SSRs in the three species genomes were compared with each other. The result showed that there were 1 410, 859, and 638 SSRs in B. japonicum, M. loti, and S. meliloti genomes, respectively. In the genomes of B. japonicum, M. loti, and S. meliloti, tetranucleotide, pentanucleotide, and hexanucleotide repeats were more abundant and indicated higher mutation rates in these species. The least abundance was mononucleotide repeat. The SSRs type and distribution were similar among these species.     

豌豆与根瘤共生对水分胁迫的生理响应

为了研究在水分胁迫条件下接种根瘤菌对豌豆植株的抗旱性影响,本试验以抗旱性品种‘定豌8号’和非抗旱性品种‘陇豌6号’两个豌豆品种为试验材料,在充分供水(田间持水量的75%)、轻度胁迫(田间持水量的55%)、重度胁迫(田间持水量的35%)3个不同水分胁迫条件下,研究了接种与不接种根瘤菌豌豆植株地上部和根系超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)、脯氨酸(Pro)、可溶性糖、丙二醛(MDA)等抗旱指标对水分胁迫的响应。结果表明:根瘤共生的豌豆植株在遭受水分胁迫后,SOD、CAT、POD活性及Pro和可溶性糖含量均高于不接菌处理,而MDA含量低于不接菌处理。接种15735菌的‘定豌8号’的3种酶活性及Pro、可溶性糖含量均高于接种Da99菌株,同时高于接种15735菌株的陇豌6号。经隶属函数综合分析表明:在3个不同的水分胁迫条件下,接种15735菌的‘定豌8号’的隶属函数平均值最大,抗旱性最强。而Da99菌株与‘陇豌6号’共生后在充分和轻度水分胁迫条件下表现出较强的抗旱性,但在重度胁迫下,Da99菌株与定豌8号表现出更强抗旱性。因此,在水分胁迫下接种两种根瘤菌均可提高豌豆的抗旱性,且15735菌对两种豌豆的抗旱性效应强于Da99菌,同时15735菌与‘定豌8号’更匹配,表现出更强的抗旱性。     

苜蓿根瘤菌固氮研究进展及浅评

综述了苜蓿根瘤菌资源调查及根瘤菌和寄主、环境间的相关特性和共生关系、共生结瘤过程与根瘤菌与寄主植物间的信息物质传递的研究,生物技术应用于根瘤菌系统发育及其分类的研究,进一步促进根瘤菌资源的开发利用等方面的研究。并对苜蓿根瘤菌固氮研究方面进行了浅评。     

几种豆科牧草根瘤的分离与接种试验

在１９８７年分离，培养４株红豆草根瘤菌（红１，红２，红３，红４）的基础上，又分离出红三叶，鹰嘴紫云英，斜茎紫花黄芪，箭舌豌豆及美国Ｎｉｔｒｏ苜蓿根瘤菌５个菌株；对已回接成功的红豆草根瘤菌株“红１”和待回接菌株“红２”及红三叶，美国Ｎｉｔｒｏ苜蓿根瘤菌进行植株感染试验。其中“红１”菌株感染力有所降低，“红２”，“红三叶”及Ｎｉｔｒｏ苜蓿菌株感染植株均获成功。从感染后所结根瘤的形态解剖特征看，均属有效     

酸、铝和盐胁迫对夏季豆科绿肥作物种子萌发及根瘤菌抗氧化酶活性的影响

在逆境土壤,种子的成功萌发与根瘤的形成是确保豆科作物正常生长的关键。本试验选择4种夏季豆科绿肥作物,包括竹豆、绿豆、田菁和豇豆,研究了种子萌发及其根瘤菌抗氧化酶活性(SOD,超氧化物歧化酶;POD,过氧化物酶;CAT,过氧化氢酶;GR,谷胱甘肽还原酶)对逆境胁迫的响应机制。种子萌发设置4个pH水平,即4,5,6,7;4个 NaCl胁迫水平,即0、40、120和200 mmol·L^-1;4个活性铝(Al³⁺)水平,即0、50、100和200 mg·L^-1。根瘤菌培养设置4个pH水平,即4,5,6,7;5个NaCl水平,即0、10、20、30和40 g·L^-1;5个活性铝(Al³⁺)水平,即0、25、50、75和100 μmol·L^-1。结果表明,酸、铝、盐胁迫显著影响几种豆科绿肥作物的种子发芽率及其根瘤菌的抗氧化酶活性,其中根瘤菌抗氧化酶活性较发芽率对胁迫敏感。不同夏季豆科绿肥品种中,田菁不耐酸,但对盐胁迫有一定耐受性;竹豆、绿豆和豇豆对低浓度的盐和弱酸性环境有一定耐受性;但是所有品种对活性铝浓度增加较为敏感。在酸性和盐碱化土壤,较高浓度的H⁺、活性铝或NaCl抑制种子萌发,减缓根瘤菌的生长,降低根瘤菌对有害物质的抗氧化能力,最终限制豆科绿肥作物的早期生长。     

rRNA基因间隔区序列用于亲缘关系密切的根瘤菌种群鉴定及系统发育分析

通过rRNA基因内转录间隔区的碱基序列分析,对中华根瘤菌属Sinorhizobium、慢生根瘤菌属Bradyrhizobium和中慢生根瘤菌属Mesorhizobium种间亲缘关系十分密切的菌株进行区分．结果表明,rRNA基因间隔区序列能很好地区分种间亲缘关系十分密切的菌株．用rRNA基因间隔区序列构建的系统发育树与rRNA基因序列构建的系统发育树结果十分相似．     

水控下大豆秸秆还田对连作土壤微生物的 影响

以连作5 a 的温室黄瓜土壤为基质，采用盆栽的方法，设置大豆秸秆不还田（CK）、接种根瘤菌大豆秸秆还田（T1）、
接种根瘤菌大豆去根瘤后秸秆还田（T2）、非接种根瘤菌大豆秸秆还田（T3）4 个处理，在不同田间持水量条件下，探究其
对土壤微生物数量的影响。结果表明，25％、50％水控条件下培养50～140 d ，T1 处理的土壤真菌数量显著低于CK，细菌
数量则在培养前期（＜ 50 d）维持较高的水平；75％水控条件下培养110、140 d 时还田处理的放线菌数量显著高于CK。总之，
25％、50％水控条件有利于维持较高的土壤细菌数量，降低真菌数量，还田处理优于非还田处理，接种处理优于非接种处理，
75％水控条件有利于培养后期（＞ 110 d）维持较高的放线菌数量。