水稻品种化感潜力的双重评价与筛选

以初筛获得的7份水稻品种为材料,用特征次生物质标记法在HPLC上测定化感指数AI值并结合田间小区试验进行双重评价与筛选,得到3份既具高化感指数又具较高田间抗草活性的水稻化感品种(系),它们是:我国台湾品种I-Kung-Pao、Parahainakoru和大陆品种HB-1。它们对无芒稗根长抑制率分别达57%、64%、55%,均超过50%;其化感指数分别是0.61、0.56、0.59,均与美国化感潜力品种PI312777的化感指数0.59相近;结果表明,水稻品种I-Kung-Pao、Parahainakoru和HB-1是我国宝贵的化感品种资源。     

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.     

旱直播条件下强弱化感潜力水稻根际微生物的群落结构

运用末端限制性片段长度多态性(T-RFLP)技术,以强化感水稻品种PI312777和弱化感水稻品种Lemont为材料,研究了旱直播模式下5叶期和7叶期化感水稻根际土壤微生物的组成和结构。结果显示:HaeⅢ和MspⅠ2种内切酶分别检测到9—87个TRFs、15—89个TRFs,各个样本的TRFs排序间存在极显著差异,表明各样本的微生物的相对丰富度存在极显著差异。用物种丰富度(S)、香隆指数(H)、Simpson多样性指数(D)、均匀度指数(E)评价了6个样本中TRFs的多样性,香隆指数最能体现2个酶切片断中TRFs的多样性,PI7中TRFs多样性最高。聚类分析和主成分分析表明旱作条件下强弱化感潜力水稻在生育前期(5叶期以前)根际微生物群落的组成与结构相似性较大,而7叶期强弱化感水稻及对照之间微生物群落差异较大,表明水稻根际微生物的组成既受生育期的影响又受水稻品种的影响。7叶期的化感水稻根际微生物呈现最大物种丰富度和香隆-威尔指数多样性,在其特有的34个菌中有7个属于粘细菌目,另外还有一些滑行菌和一些与氮循环相关的细菌等。粘细菌可能通过次生代谢产物抑制伴生杂草种子的萌发,降低杂草的密度从而提高化感水稻的化感作用潜力。     

Isolation and characterization of a potential biocontrol agent Bacillus KM5 from rhizosphere soil of a rice plant

Twelve microbial isolates were isolated and purified from rhizosphere soil of a healthy rice plant. One of the isolate named as KM 5 showed antagonist activity in vitro against Sclerotium rolfsii Saccardo, Helminthosporium oryzae, Gibberella fujikuroi, Rhizoctonia solani Nees and Fusarium udum. KM 5 was characterized by microscopic, Gram stain and biochemical methods belonging to genus Bacillus. The genus Bacillus was further confirmed by isolation of genomic DNA, 16S rRNA amplification using Bacillus specific primers. Sequencing of the PCR amplified product using 775 base pairs and further NJ phylogenetic analysis confirmed that the microorganism is a new strain of bacterium named as Bacillus sp. KM 5. Partial sequence of 16S rRNA gene has been deposited to NCBI GenBank data base with accession no. EU266068 and deposited to MTCC with accession no. MTCC-5413. The cell free culture filtrate of Bacillus sp. KM 5 inhibited the growth of all test fungi. An antifungal protein produced by Bacillus sp. KM 5 was purified by ammonium sulphate. The protein was stable at 20, 40, 60 and 100°C and remained active after sterilization at 121°C for 15 min.     

水稻品种化感特性与农艺性状的关系

选用9个华南地区主栽水稻品种,用特征次生物质标记方法测定它们的化感指数,并通过田间实验确定这些水稻品种的抑草效应及主要农艺性状,在此基础上分析这些水稻品种的化感特性、抑草效应和农艺性状的关系.结果表明,具有化感特性的水稻品种并不都能在田间表现出抑草效应,化感特性与主要农艺性状均无显著的相关性,但同时具有化感特性和有利竞争农艺性状的水稻品种均能在田间显示抑草效应.水稻化感特性和农艺性状的不相关显示化感特性转入相关的水稻品种将不影响它们的农艺性状,这意味着能培育出各种农艺性状的水稻化感新品种.     

植物化感物质及化感潜力与土壤养分的相互影响

植物化感作用与许多生态因子有关.土壤养分缺乏,影响着许多植物化感物质的产生,从而影响植物的化感潜力;反过来,植物化感物质也通过络合、吸附、酸溶解、竞争、抑制等方式影响土壤的养分形态和水平.本文总结了植物化感物质及化感潜力与土壤养分的相互影响,并提出了今后该领域值得进一步研究的问题.包括以下几方面：加强植物化感研究与土壤 植物营养学研究的结合,以更深入地阐明植物化感物质、化感作用与土壤养分变化的关系;加强植物化感研究与生态系统养分循环研究的结合,以类似自然（nature-like）的方式模拟自然界植物所受的养分干,使养分干扰的化感研究结果更加逼真、可靠;加强对养分过量及受污染时植物化感作用的研究,为揭示农业和林业生产中植物的相互作用机制和生物量变化机制提供新的思路,为生态保护提供科学依据.     

不同施肥处理下水稻根际和非根际土壤中氨基糖积累特征

以水稻长期定位施肥试验土壤为研究对象,选取不施肥(CK)、化肥(NPK)、秸秆还田+化肥(NPKS)、30%有机肥+70%化肥(LOM)和60%有机肥+40%化肥(HOM)5种处理,分析水稻分蘖旺期根际土和非根际土中氨基糖积累特征.结果表明: 与CK和NPK处理相比,长期施用有机物料(NPKS、LOM、HOM)显著增加了水稻根际土和非根际土中有机碳、总氨基糖及其氨基单糖(胞壁酸、氨基葡萄糖和氨基半乳糖)含量.不同施肥处理下3种氨基单糖的积累规律不同,说明不同微生物对施肥处理的响应趋势和强度有所不同.受稻田翻耕等均匀化土壤的农事操作影响,各处理总氨基糖含量在根际土与非根际土间无显著差异.氨基糖碳对土壤有机碳积累的贡献范围为24.0～28.3 mg·g^-1,且以NPKS处理最高,HOM和CK处理最低.真菌氨基葡萄糖/胞壁酸比值范围为24.4～36.6,说明该试验点所有处理的根际土与非根际土中有机质的降解与转化过程以真菌为主导,且与NPK和CK相比,NPKS处理的真菌参与度提高,而施用HOM处理的细菌参与度提高.     

Molecular,biochemical and bioassay based evidence of lower allelopathic potential in genetically modified rice

This study examines allelopathic potential of genetically modified rice. The experiment was conducted on two isogenic lines Bacillus thuringiensis (Bt) and non-Bacillus thuringiensis (non-Bt). Both isogenic lines have same allelopathic ability before insect feeding and after limited insect feeding (Spodoptera litura) non-Bt rice genotype demonstrates more allelopathic potential. The S. litura cannot feed Bt rice genotype. The role of shoot herbivory in allelopathic induction is further supported when Bt plants also exhibited higher allelopathic potential after insect regurgitant application to the damaged leaves. Allelopathic potential was assessed through several methods after treatments of mechanical damage, insect feeding and insect regurgitant application to damaged rice leaves. Rhizosphere soil and leaf leachates of non-Bt rice cultivar exhibited higher allelopathic potential on lettuce and barnyard grass after herbivore feeding. Enzyme activities (PAL and C4H) responsible for biosynthesis of phenolic compounds and their concentration were significantly higher in non-Bt plant after herbivore feeding and attain the same level in Bt plants after insect regurgitant application to damaged leaves. Similarly, genes (OsPAL and OsCYC1) responsible for biosynthesis of allelopathic compounds showed high expression in non-Bt plants after herbivore feeding. Our results indicate that herbivore feeding enhance rice allelopathic potential and no insect feeding as incase of Bt plants may reduce allelopathic potential of genetically modified rice.     

低钾胁迫对水稻(Oryza sativa L.)化感潜力变化的影响

研究以国际公认的化感水稻P1312777和非化感水稻Lemont为供体,稗草（Echinochloa cru-galli L.）为受体,采用稻／稗共培体系,研究低钾胁迫对水稻化感潜力变化的影响及其机制。受体稗草的形态指标分析结果表明,低钾胁迫促使化感水稻P1312777对共培稗草的根长、株高和干重的抑制率均升高,增幅远大于非化感水稻Lemont。受体稗草生理生化指标分析结果表明,低钾胁迫下化感与非化感水稻对受体稗草保护酶系（SOD、POD、CAT）及根系活力的抑制作用增强,但化感水稻P1312777比非化感水稻Lemont的抑制程度大,且达极显著差异。实时荧光定量PCR分析结果表明,低钾胁迫下,化感水稻P1312777根部与叶部中酚类代谢的关键酶——苯丙氨酸解氨酶、肉桂酸-4-羟化酶、羟化酶、O-甲基转移酶的基因均上调表达,而非化感水稻根部相应酶均下调表达,叶部除苯丙氨酸解氨酶上调,其余酶也下调表达。而萜类代谢途径关键酶——HMG—CoA还原酶、角鲨烯合酶、单萜烯环化酶、倍半萜烯环化酶、二萜烯环化酶的基因,在两种水稻根部中呈现出相同或相似的表达方式（上调或下调）,即HMG—CoA还原酶上调表达,角鲨烯合酶、单萜烯环化酶、倍半萜烯环化酶、二萜烯环化酶下调表达;而在水稻叶部,非化感水稻Lmont相应酶基因表达方式仍然不变,化感水稻P1312777除了角鲨烯合酶下调表达,其余4个酶均上调表达。水稻根系分泌物中酚类物质的HPLC分析结果表明,低钾胁迫下,化感水稻P1312777根系分泌物中,所检出的酚酸类物质总量是正常营养条件下的2．30倍,而非化感水稻Lemont则是正常营养条件下的0．91倍。综合分析认为低钾胁迫下,化感水稻P1312777抑草能力增强主要是由于酚类代谢途径关键酶基因表达上调,导致酚类代谢途径旺盛,分泌出更多的酚类物质,进而破坏受体稗草保护酶系统,抑制了稗草的正常生长。     

The allelopathic potential of alfalfa root medicagenic acid glycosides and their fate in soil environments

Summary The allelopathic effect of alfalfa (Medicago media Pers.) and red clover (Trifolium pratense L.) root saponins on winter wheat seedling growth and the fate of these chemicals in soil environments were studied. Seed germination, seedling and test fungus growth were suppressed by water and by alcohol extracts of alfalfa roots, and by crude saponins of alfalfa roots, indicating that medicagenic acid glycosides are the inhibitor. Powdered alfalfa roots inhibited wheat seedling growth when added to sand. At concentrations as low as 0.25% (w/w) the root system was completely destroyed whereas seedling shoots suffered little damage. Red clover roots caused some wheat growth inhibition when incorporated to sand, but their effect was much lower than in the alfalfa root treatment. Soil textures had a significant influence on the inhibitory effect of alfalfa roots. The inhibition of seedling growth was more pronounced on light than on heavy soils. This was attribted to the higher sorption of inhibitors by heavy soils. Incubation of alfalfa roots mixed into loose sand, coarse sand, loamy sand and clay loam for a period of 0–8 days resulted in decreased toxicity to bothT. viride and wheat seedlings. This decrease occurred more quickly in heavier soils than in loose sand, due to the hydrolysis of glycosides by soil microorganisms. Soil microbes were capable of detoxifying medicagenic acid glycosides by partial hydrolysis of sugar chain to aglycone. These findings illustrate the importance of medicagenic acid glycosides as an inhibitor of wheat seedling growth, and of their fate in different soil environments.     

Stress-induced allelopathic activity and momilactone B in rice

Allelopathic activity of rice extracts and root exudates against Echinochloa crus-galli increased by heavy metal, cantharidin and jasmonic acid treatments. Since cantharidin (protein phosphatase inhibitor) acts as an elicitor and jasmonic acid is an important signaling molecule regulating inducible defense genes against the pathogen infection, heavy metal stress and pathogen infection may increase alleopathic activity of rice. These treatments also increased the concentrations of momilactone B in rice extracts and root exudates, suggesting that the production of momilactone B in rice and the secretion of momilactone B from rice into the rhizosphere may be enhanced by the treatments. As momilactone B possesses strong phytotoxic and allelopathic activities, the elevated production and secretion of momilactone B of rice by heavy metals, cantharidin and jasmonic acid may contribute to the increasing allelopathic activity of rice. Enhancement of the secretion of momilactone B into the rhizosphere may provide a competitive advantage for root establishment through local suppression of pathogen and inhibition of the growth of competing plant species. Therefore, allelopathy of rice may be one of the inducible defense mechanisms and may be regulated several environmental factors.     

Conditional allelopathic potential of temperate lianas

Allelopathy is often treated as an innate characteristic of a species rather than a phenotypically plastic trait that can vary with environmental conditions. Lianas are a highly competitive, phenotypically plastic life form that typically occur in both shaded and unshaded environments. As such, we hypothesized that temperate lianas may conditionally change allocation to allelopathic chemicals in response to light availability though the expected direction of change is unclear. Shading may reduce resource availability and therefore reduce allocation to allelochemicals, induce allelopathy as a competitive mechanism, or may not be related to allelopathy. To test the conditionality of allelopathy, sun and shade leaves of five common liana species (Toxicodendron radicans, Parthenocissus quinquefolia, Celastrus orbiculatus, Lonicera japonica, and Vitis vulpina) were collected from a young deciduous forest in New Jersey, USA, and tested with laboratory bioassays to detect allelopathic potential. All liana species showed allelopathic potential, and three species exhibited induction of increased allelopathic potential in shaded environments. The two species that were not shade induced are late successional lianas that persist for long periods in forest canopies. In contrast, the inducible lianas were early successional species that typically decline with canopy closure. This research indicates that lianas have the potential to be allelopathic and allelopathic potential conditionally responds to shading only for species that would normally be excluded from the forest canopy. As early successional lianas are present throughout forest regeneration in a range of light environments, allelopathic plasticity may increase their success by differentially allocating resources based on environmental conditions.     

不同化感水稻品种根系分泌物中萜类化合物的差异分析

以强化感水稻品种PI312777和弱化感水稻品种Lemont为供试材料,运用“循环法”收集供试水稻经土壤和石英沙介质后的根系分泌物,并经乙醚萃取,所获萃取物经气相色谱 质谱联用仪（GC-MS）测试,检索谱图库确定化合物.结果表明,乙醚萃取物中检测到含氧萜类化合物,如柠檬烯氧化物、柠檬烯二氧化物、香芹酮氧化物、香芹醇、雪松醇等,不同品种间含氧萜类化合物种类相似,但在不同介质中的存留方式和数量存在差异.文中还就不同培养方式和品种间萜类化合物的差异作了进一步讨论.     

菊科植物化感作用研究进展

对菊科植物化感作用的研究进展进行了综述。菊科植物中至少有 39个属存在化感作用 ,特别是一枝黄花属、向日葵属、胜红蓟属、银胶菊属、蒿属植物等有较多的研究报道。鉴定出的化感物质多为萜类、聚乙炔类、酚类、有机酸类等 ,这些化感物质对多种受体植物表现出程度不同的抑制或促进的效应。其化感作用机理表现在破坏受体膜系统的稳定性及水分平衡关系、抑制氧化磷酸化、促进或阻滞叶绿素的合成、影响矿质元素的吸收利用等。并对菊科植物化感物质在植物生长调节剂、天然除草剂和生物杀虫剂 ,或人工合成除草剂和杀虫剂上应用的前景进行了探讨。本文显示菊科植物的化感作用将在控制外来恶性杂草及维护生态平衡上扮演重要的角色。在当前菊科植物化感作用研究的基础上 ,提出了进一步研究的 6个方向 :(1)化感物质的生物合成途径与关键酶的特性研究 ;(2 )具化感潜势物种资源的调查评价及利用研究 ;(3)化感作用在自然生态系统中的演变规律 ;(4 )菊科重要作物自毒的生化机制及克服途径 ;(5 )具应用前景的菊科植物化感关键酶的基因克隆和转基因 ,并对受体植物基因的表达与调控进行研究 ;(6 )化感作用在可持续发展农业应用上的研究与开发 ,特别是作为天然除草剂及杀虫剂     

水稻化感品种对土壤微生物的影响

通过盆栽实验研究三叶期水稻化感品种对土壤微生物种群数量的影响。结果显示,土壤细菌、放线菌、真菌、氨化细菌和好气性自生固氮菌数量在水稻化感品种P1312777和水稻非化感品种“辽粳九”、“秋光”的土壤中存在显著差异,土壤中大多数微生物被水稻化感品种所抑制,但化感品种土壤中的放线菌和好气性纤维素分解菌数量则介于两种非化感品种之间。这一研究表明,水稻化感品种能显著地影响土壤微生物种群数量。     

基于根际与凋落物际评价转Bt水稻对土壤线虫群落的影响

转基因作物在商业化推广前,有必要评价其对非靶标土壤生物的生态影响。根系和凋落物与土壤的界面是代表植物直接影响土壤的两个典型活性微域,即根际和凋落物际。基于室内盆栽实验构建水稻根际和凋落物际,比较了这两个高活性微域中转Bt水稻克螟稻(KMD)与亲本水稻秀水11(XSD)对土壤可利用资源、微生物学性质和线虫群落的影响。结果表明:转Bt水稻对土壤性质的影响依赖于水稻生育期和微域的变化。与XSD相比,KMD在苗期和拔节期显著提高了凋落物际土壤可溶性有机氮含量(P0.05);在苗期显著降低了凋落物际土壤铵态氮含量,但在成熟期显著提高了凋落物际与交互微域土壤的铵态氮含量(P0.05)。与亲本水稻相比,转Bt水稻分别在苗期和成熟期显著提高了凋落物际土壤微生物生物量碳和活性,而在拔节期和成熟期显著降低了根际土壤微生物生物量氮含量(P0.05)。微域间的交互作用显著影响土壤微生物生物量、可溶性有机碳和氮、线虫总数和各食性线虫的数量(P0.05)。转Bt水稻总体上降低了土壤线虫总数,尤其在苗期和拔节期的凋落物际土壤中达到显著水平。重复测量方差分析表明转Bt水稻显著影响土壤食细菌(P0.01)和食真菌(P0.05)线虫数量,及线虫群落的通道指数和富集指数。水稻收获后KMD秸秆的Bt蛋白含量显著高于亲本,全部KMD处理的土壤样品中Bt蛋白含量无显著变化,因而转Bt水稻对土壤生物学性质的影响可能并非来源于Bt蛋白的作用,而更可能来自于水稻生长性状和凋落物性质的差异。