分蘖洋葱根系分泌物对黄瓜幼苗生长及根际土壤微生物的影响

以不同化感潜力分蘖洋葱为供体,黄瓜为受体,研究了分蘖洋葱根系分泌物对黄瓜幼苗生长、根际土壤微生物数量及细菌群落结构的影响.结果表明:不同化感潜力分蘖洋葱根系分泌物对黄瓜幼苗生长均具有促进作用,且随着浓度的升高,促进作用增强,相同浓度下,化感潜力强、弱供体之间差异不显著;不同化感潜力分蘖洋葱根系分泌物均增加了黄瓜根际土壤细菌和放线菌数量,降低了真菌和尖镰孢菌数量,化感潜力强的品种(L-06)效果更显著;不同化感潜力分蘖洋葱根系分泌物均能提高黄瓜根际土壤细菌群落丰富度,差异条带的序列片段经比对推测为3大细菌类群:Actinobacteria(放线菌纲)、Proteobacteria(变形菌纲)和Anaerolineaceae(厌氧绳菌纲),其中厌氧绳菌只出现在化感潜力强(L-06)的处理中.化感潜力强(L-06)、浓度高(10 mL·株-1)的分蘖洋葱根系分泌物更有利于黄瓜根际土壤细菌群落丰富度的提高.     

轮套作对黄瓜根际土壤细菌种群的影响

以黄瓜为试材,采用PCR-DGGE技术研究了小麦、大豆、毛苕子、三叶草和苜蓿与黄瓜轮作及大蒜、毛葱与黄瓜套作对黄瓜根际土壤细菌种群的影响.结果表明:轮套作对黄瓜根际土壤细菌种群的种类、数量和黄瓜产量具有一定的影响,提高了土壤细菌种群多样性指数、均匀度指数和黄瓜产量.对DGGE条带进行测序的结果表明,它们大多与不可培养的细菌种属具有较高的同源性,大多为鞘氨醇杆菌属（Sphingobacterium）和变形细菌纲（Proteobacteria）, High bacterium G+C只在以毛葱为套作作物的黄瓜根际中出现.不同轮套作中细菌种群多样性随黄瓜生育期的变化而变化,在盛瓜期达最高值.以小麦为轮作作物和以毛葱为套作作物的黄瓜轮套作栽培模式最佳.     

小麦根系分泌物对黄瓜生长及土壤真菌群落结构的影响

以黄瓜为受体,以不同化感效应(促进/抑制)小麦品种为供体,采用PCR-DGGE技术,研究了小麦根系分泌物及伴生小麦对黄瓜生长及土壤真菌群落结构的影响.结果表明： 在处理第6天和第12天,化感促进效应小麦根系分泌物分别显著提高了黄瓜幼苗株高和茎粗;在处理第18天,化感促进和抑制效应小麦根系分泌物均显著提高了黄瓜幼苗株高;在处理第6天,不同化感效应小麦根系分泌物均显著降低了黄瓜幼苗根际土壤真菌群落条带数、Shannon指数及均匀度指数,有苗对照(W)显著高于无苗对照(Wn);在处理第18天,各处理的真菌群落结构条带数、Shannon指数及均匀度指数均显著高于无苗对照(Wn).伴生化感抑制效应小麦显著降低了黄瓜根际土壤真菌群落Shannon指数和均匀度指数,说明小麦根系分泌物及伴生小麦改变了土壤真菌群落结构.DGGE图谱及其主成分分析结果表明,伴生不同化感效应小麦对土壤真菌群落结构影响较大.     

国家自然科学基金项目简介

<正>项目名称:分蘖洋葱PAL和ALL基因调控主效化感物质促黄瓜生长的机理研究项目经费:2 3万项目主要内容:套作分蘖洋葱可提高黄瓜产量,这与分蘖洋葱释放的化感物质有关,其分子调控机制尚有待研究。本项目以化感物质合成代谢途径关键酶基因PAL和ALL为切入点,采用RACE、RNAi、RNA seq等技术试图阐明PAL和ALL基因调控主效化感物质促黄瓜生长的机理。该研究结果可为揭示套作提高产量的机制,以及将分蘖洋葱化感物质开发成为植物生长调节剂     

抗草甘膦转基因大豆（RRS）对根际土壤细菌数量和多样性的影响

为深入研究种植抗草甘膦转基因大豆的黑土生态区根际土壤中细菌数量及多样性的变化,试验采用DGGE-cloning测序技术与传统培养相结合的方法,研究了抗草甘膦转基因大豆(RRS)对根际土壤细菌数量以及细菌群落多样性的影响.传统培养试验结果为RRS显著降低了土壤细菌的数量;DGGE图谱分析表明,RRS根际土壤细菌16SrDNA条带数、多样性指数及均匀度指数均要低于其他处理,聚类分析显示RRS带谱与RRS-S和Y-S差异较大,相似性分别为64%和64.4%;DGGE-cloning测序结果表明,在RRS处理中缺失条带1和条带12分别属于Uncultured bacterium和Nitrospira门Nitrospira属,其中条带1与其他切取条带最小遗传距离达0.4,与其他处理相比表现出弱势差异的条带2、4、5和条带11均属于Uncultured bacterium.研究表明,RRS不同程度上降低了根际土壤细菌的数量和细菌群落的多样性,并对根际土壤中Nitrospira属细菌有一定的抑制作用.     

福建省稻田土壤细菌群落的16S rDNA-PCR-DGGE分析

用不依赖细菌培养的16S rDNA-PCR-DGGE方法对福建省6个不同地区12个取样点的稻田土壤进行细菌群落结构分析.对12份样品直接提取其总DNA,用F341GC/R534引物扩增16SrDNA基因的V3可变区,结合DGGE(denaturing gradient gel electrophoresis)技术分析样品细菌群落组成.结果表明,福建省不同地区的稻田土壤之间细菌群落结构存在较大差异.犬体上可分为闽东、闽南、闽北、闽西4个大类.同一地区的根际土和表土样品之间也存在差异,但差异相对较低,其中龙岩根际土和表土细菌群落结构相似性最大,永泰差异性最大.回收了DGGE图谱中11个条带,测序结果经过Blast比对表明其中10个条带代表的细菌是不可培养的,显示了DGGE技术的优越性.     

化感水稻根际微生物类群及酶活性变化

以化感水稻PI312777（PI）和非化感水稻Lemont（LE）为材料,分别测定不同水稻叶龄期（3～7叶期）根际微生物区系变化及根际土壤酶活性.结果表明,化感水稻明显影响土壤根际微生物类群及相关酶活性.化感水稻PI根际细菌、放线菌、固氮菌的数量高于非化感水稻LE,增幅分别在11.2%～28.3%、40%～78.6%和111.5%～173.9%之间,而真菌数量低于非化感水稻LE,最高仅为其值的25.5％,说明化感水稻PI对绝大多数细菌、放线菌、固氮菌生长有促进作用,对一些真菌生长有抑制作用.进一步分析表明,化感水稻PI对氨化细菌、亚硝酸细菌、硝酸细菌、好气性固氮菌、好气性纤维素分解菌、硫化细菌的生长具有促进作用,其中以氨化细菌、好气性固氮菌的更为明显,最低增幅分别为53.7％和57.6％;而对反硫化细菌、反硝化细菌生长有抑制作用,其值最高分别为非化感水稻的54.2％和50.6％.此外,化感水稻PI根系分泌物对脲酶、磷酸酶、蔗糖酶的活性具有促进作用,而对过氧化氢酶则呈抑制作用.     

盐碱胁迫对巨菌草根际土壤微生物多样性及酶活性的影响

为探明盐碱胁迫对巨菌草根际土壤微生物多样性及酶活性的影响,该文设置了7个盐碱梯度,利用SPSS和Illumina高通量测序分析巨菌草根际土壤生理指标和微生物多样性.结果表明:(1)各处理间,巨菌草根际土壤真菌和细菌结构均存在差异;12‰盐碱浓度下,真菌优势菌纲为粪壳菌纲(68.5％)、散囊菌纲(16.3％),细菌优势菌...     

RISA、DGGE和2D-PAGE技术对大豆根际土壤细菌多样性分析的比较

采用基于PCR扩增的核糖体间隔区分析(RISA)、变性梯度凝胶电泳(DGGE)和双向电泳(2D-PAGE)3种分子生态学技术对大豆根际土壤细菌多样性比对分析.结果表明:2D-PAGE技术得到的土壤细菌多样性(基因点)最丰富,其次为DGGE技术(基因片段),RISA技术(基因片段)最低.RISA技术得到的条带数最少,但结...     

嫁接对铜胁迫下黄瓜根际土壤微生物特性和酶活性的影响

采用盆栽试验方法,研究了嫁接（以黑籽南瓜为砧木）对铜胁迫下黄瓜根际土壤微生物生物量、微生物种群数量和土壤酶活性的影响.结果表明： 铜胁迫下黄瓜根际土壤微生物生物量碳（MBC）和微生物生物量氮（MBN）含量显著下降,基础呼吸和代谢熵显著上升,但嫁接黄瓜根际土壤MBC和MBN含量显著高于自根黄瓜,而基础呼吸和代谢熵则显著低于自根黄瓜.铜胁迫下,根际土壤放线菌和自生固氮菌的数量显著下降,真菌数量显著上升,而细菌数量变化不显著;嫁接黄瓜根系土壤细菌、放线菌、自生固氮菌的数量显著高于自根黄瓜,而真菌数量显著低于自根黄瓜.嫁接黄瓜根际土壤脲酶、磷酸酶、蔗糖酶、过氧化氢酶活性在铜胁迫下显著高于自根黄瓜.试验结果证明嫁接使铜胁迫下黄瓜根际土壤微生物环境和酶活性得到了改善和提高,从而提高了黄瓜植株对铜胁迫的抵抗力.     

Effects of intercropping Chinese onion cultivars of different allelopathic potential on cucumber growth and soil micro-environment

A pot experiment was conducted to study the effects of intercropping various Chinese onion cultivars of different allelopathic potential on the cucumber growth and rhizospheric soil environment. When intercropped with high allelopathic Chinese onion cultivars, the EC value and peroxidase activity of cucumber rhizospheric soil decreased, while the pH value, invertase and catalase activities, and bacterial community diversity increased. The cloning and sequencing results indicated that most DGGE bands amplified from cucumber rhizospheric soil samples showed a high homology to uncultured bacterial species. The common bands were affiliated with Actinobacteria and Proteobacteria, and the differential bacteria bands were affiliated with Proteobacteria and Anaerolineaceae. Rhodospirillales and Acidobacteria were only found in the cucumber rhizospheric soil intercropped with low allelopathic Chinese onion cultivars. Correlation analysis showed that there were significant positive correlations between rhizospheric soil urease activity and cucumber seedlings height, total dry biomass, leaf area, and DGGE band number. It was suggested that intercropping high allelopathic Chinese onion cultivars could establish a good rhizospheric soil micro-environment for cucumber growth, and promote the growth of cucumber seedlings markedly.     

Impact of allelopathic rice seedlings on rhizospheric microbial populations and their functional diversity

Field performance of rice allelopathic potential is indirectly regulated by the microflora in the rhizosphere. The present study aimed to investigate the dynamics of microbial populations and their functional diversities in the seedling rhizospheres of rice cultivars with varied allelopathic activities by employing agar plate bioassay, fumigation and BIOLOG analysis. Rice cultivars significantly affected the microbial carbon content in their associated rhizospheric soil. The microbial carbon contents were ranked in a decreasing order as Iguape Cateto (441.0 mg·kg^–1) > IAC47 (389.7 mg·kg^–1) > PI312777 (333.2 mg·kg^–1) > Lemont (283.8 mg·kg^–1) with the nil-rice control soil of 129.3 mg·kg^–1. Similarly, the respiration rate of the soils was 1.404, 1.019, 0.671 and 0.488 μgC·g^–1· h^–1 for PI312777, Iguape Cateto, IAC47 and Lemont, respectively. The respiration rate was only 0.304 μ gC·g^–1·h^–1 for the control soil. The microbial flora in the rhizospheric soil of different rice cultivars was dominated by bacteria (58.4%–65.6%), followed by actinomycete (32.2%–39.4%) and fungi (2.2%–2.8%). BIOLOG analysis showed that the value of Average Well Color Development (AWCD) differed significantly among rice cultivars. It was always the highest in the rhizospheric soil of the strongly allelopathic rice cv. PI312777, and the lowest in the rhizospheric soil of the poorly allelopathic rice cv. Lemont. The AWCD value reached the maximum in all the sampled soils after 144 hours of incubation. The AWCD values from the rhizospheric soils of PI312777, IAC47, Iguape Cateto and Lemont were 1.89, 1.79, 1.60 and 1.43 times higher than that of the control soil. Principal Component Analysis (PCA) identified 3 principal component factors (PCF) in relation to carbon sources, accounting for 70.1%, 11.3% and 7.0% of the variation, respectively. 19 categories of carbon sources were significantly positively correlated to the 3 principal components. Phenolic acids, carbohydrates, amino acids and amides were significantly correlated to the principal component 1, phenolic acids, carbohydrates and fatty acids to the principal component 2, and carbohydrates and hydroxylic acids to the principal component 3. Amino acids and amides were the two main carbon sources separating the 3 principal component factors. In addition, the total microbial population in the rhizospheric soil was significantly positively correlated with AWCD, microbial biomass carbon, microbial respiration and Shannon index. There was a significantly positive correlation between the total microbial population and the inhibition rate (IR) on the root length of lettuce owing to the different allelopathic activities of the rice cultivars. These results suggest that changes in microbial population, activity and functional diversity in the rhizospheres are highly cultivar-dependent. These changes might play an important role in governing the rice allelopathic activity in the field.     

Protein expression in accessions of Chinese onion with different allelopathic potentials under monocropping and intercropping systems

Protein expressions of Chinese onion accessions grown under monoculture and intercropped with cucumber were evaluated in pot experiments. Chinese onion accessions used were L04 (with weak allelopathic potential) and L06 (with strong allelopathic potential). Root proteins were separated by two-dimensional electrophoresis and the variable expressed protein spots were identified with MALDI-TOF-TOF mass spectrometer. Forty-seven identified proteins were classified into nine functional categories. Compared monocropping and intercropping, 31 identified variable protein spots were classified into energy metabolism (14 %), phenylpropanoid biosynthesis (28 %), organosulfur compounds biosynthesis (25 %), carbohydrate metabolism (10 %), fatty acid hydrogen peroxide metabolism (9 %), protein translation (3 %), other function (3 %), and no assigned function (9 %). Compared Chinese onion accessions of differing allelopathy potentials, 22 identified variable protein spots were classified into energy metabolism (18 %), phenylpropanoid biosynthesis (27 %), organosulfur compounds biosynthesis (23 %), carbohydrate metabolism (9 %), nucleosome component (4 %), other function (14 %), and no assigned function (5 %). The level of variable-expressed proteins involved in phenylpropanoid and organosulfur compounds biosynthesis significantly upregulated in treatments intercropped with cucumber. These results suggested that putative allelochemicals of Chinese onion were mainly produced by phenylpropanoid biosynthesis and organosulfur compounds biosynthesis pathway.     

Positive allelopathic stimulation and underlying molecular mechanism of achyranthe under continuous monoculture

Three experiments were conducted to study the allelopathic stimulation and its underlying molecular mechanism of achyranthes medicinal plants in continuously monoculture system. The stimulators in the rhizospheric soil of continuously monocultured achyranthes plants were extracted by water and organic solvents. Results of the bioassay showed that the rhizospheric soil extracts had a significant promotive effect on the growth of achyranthes in continuous monoculture system, implying that the extracts, especially the water extracts might contain plant activators to stimulate the growth of the medicinal plants. Subtractive hybridization suppression (SSH) was used to investigate gene expression profiles of achyranthes in response to the extract treatments. Ten up-regulated genes from SSH-cDNA library were sequenced and assigned. Results indicated that flavonoids and phytosterol might play an important role in the positively allelopathic stimulation on achyranthes plants in continuous monoculture system. Comparative proteomics were employed to further unveil the molecular mechanism of allelopathic stimulation induced by the extracts. Compared with protein expression profile in control, 25 differentially expressed proteins and their functions were detected and identified in the treated plants. The results suggested that the extracts from continuously monocultured rhizospheric soils under Chinese medicinal achyranthes activated the genes encoding the key enzymes involved in terpenes and flavonoids synthesis, which in turn led to increased de novo synthesis of the stimulators, and hence promoted growth of achyranthes in consecutively monoculture system.     

Crotonic acid as a bioactive factor in carrot seeds (Daucus carota L.)

Water extracts from the carrot seed (Daucus carota L.) var. Perfekcja exhibit plant growth inhibitory properties against cress, cucumber, onion and carrot in a dose-dependant manner. This property results from the action of low-and high-molecular components of the extract. The low-molecular component was identified as crotonic acid ((E)-2-butenoic acid). Its presence was also confirmed in other late varieties of carrot. The determined strong herbicidal properties of crotonic acid and its availability after release to soil combined with its high level in seeds suggest that it might be considered as an allelopathic and autotoxic factor in the seeds.     

Effects of root-knot nematodes on cucumber leaf N and P contents, soil pH, and soil enzyme activities

A pot experiment was conducted to study the effects of inoculation with root-knot nematodes on the cucumber leaf N and P contents, and the rhizospheric and non-rhizospheric soil pH and enzyme activities. The rhizospheric soil pH didn't have a significant decrease until the inoculation rate reached 6000 eggs per plant. With the increase of inoculation rate, the leaf N and P contents, rhizospheric soil peroxidase activity, and rhizospheric and non-rhizospheric soil polyphenol oxidase activity all decreased gradually, rhizospheric soil catalase activity was in adverse, non-rhizospheric soil pH decreased after an initial increase, and non-rhizospheric soil catalase activity had no regular change. After inoculation, rhizospheric soil urease activity decreased significantly, but rhizospheric and non-rhizospheric soil phosphatase activity and non-rhizospheric soil peroxidase activity only had a significant decrease under high inoculation rate. In most cases, there existed significant correlations between rhizospheric soil pH, enzyme activities, and leaf N and P contents; and in some cases, there existed significant correlations between non-rhizospheric soil pH, enzyme activities, and leaf N and P contents.     

Comparing Anthracene and Fluorene Degradation in Anthracene and Fluorene-Contaminated Soil by Single and Mixed Plant Cultivation

The ability of three plant species (sweet corn, cucumber, and winged bean) to remediate soil spiked with 138.9 and 95.9 mg of anthracene and fluorene per kg of dry soil, respectively, by single and double plant co-cultivation was investigated. After 15 and 30 days of transplantation, plant elongation, plant weight, chlorophyll content, and the content of each PAH in soil and plant tissues were determined. Based on PAH removal and plant health, winged bean was the most effective plant for phytoremediation when grown alone; percentage of fluorene and anthracene remaining in the rhizospheric soil after 30 days were 7.8% and 24.2%, respectively. The most effective combination of plants for phytoremediation was corn and winged bean; on day 30, amounts of fluorene and anthracene remaining in the winged bean rhizospheric soil were 3.4% and 14.3%, respectively; amounts of fluorene and anthracene remaining in the sweet corn rhizospheric soil were 4.1% and 8.8%, respectively. Co-cultivation of sweet corn and cucumber could remove fluorene to a higher extent than anthracene from soil within 15 days, but these plants did not survive and died before day 30. The amounts of fluorene remaining in the rhizospheric soil of corn and cucumber were only 14% and 17.3%, respectively, on day 15. No PAHs were detected in plant tissues. This suggests that phytostimulation of microbial degradation in the rhizosphere was most likely the mechanism by which the PAHs were removed from the spiked soil. The results show that co-cultivation of plants has merit in the phytoremediation of PAH-spiked soil.