相同遗传背景不同植株形态水稻等基因系与杂草的竞争及化感作用

利用相同遗传背景不同株形水稻浙辐802等基因系为材料,研究了IG1、IG4、IG25和IG26与无芒稗的竞争和化感作用潜力,探索了二者之间的相互关系.结果表明,IG1和IG4的植株高度和分蘖角度显著大于IG25和IG26,但它们的根系活力却显著小于IG25和IG26.IG1和IG4对无芒稗生物量干重抑制率分别达到15%和17%,显著高于IG26的64%,表明水稻与无芒稗竞争力与水稻植株高度和分蘖角度成正比,而与水稻根系活力成反比.IG1叶片水浸提液对无芒稗根长抑制率为77.6%,显著高于非化感对照品种秀水63,而植株形态相近的IG4的化感作用却未达到显著水平,显示在相同遗传背景下水稻化感作用强弱与水稻株型无相关性.     

不同类型稻种资源对稗草化感潜力差异评价

以120份不同类型稻种资源为研究对象,将田间鉴定和室内叶浸提液处理鉴定方法相结合,对其化感作用差异进行了鉴定评价.结果表明,在田间不同类型稻种资源对稗草生长的抑制效果大小趋势为地方品种＞选育品种(系)＞引进品种(系)＞杂交水稻(恢复系和不育系),对稗草苗鲜重和苗干重的抑制效果大于对苗高的抑制效果.室内叶浸提液处理鉴定结果,不同类型稻种资源间化感作用差异相对较小,但仍以地方品种较强,并对稗草根长的抑制效果大于对稗草苗高的抑制效果.地方品种中具有较强化感作用的种质资源频率高于其他类型品种,今后要加强对地方品种资源的化感潜力鉴定和评价.     

化感作用水稻对无芒稗及相关田间杂草的抑制作用评价

对从500份水稻种质资源中用差时播种共培法的改进方法初步筛选出10份水稻化感抑稗材料进行了田间化感抑草评价研究.结果表明,在直播条件下,化感材料谷梅2号、吓一跳、鸡早籼、赣早籼和商糯1号等对无芒稗的抑制作用显著优于水稻无化感作用材料秀水63和春江11.化感材料Milyang54对田间总量杂草的抑制作用最强.在移栽条件下,化感材料谷梅2号、青困2号、吓一跳和鸡早籼对无芒稗的抑制作用与无化感作用材料秀水63和春江11差异及显著性水平.水稻材料间部分农艺性状差异及显著性水平,化感材料吓一跳和鸡早籼的株高较高,分蘖力较强,但顶三叶叶面积较小.     

不同供氮水平下反枝苋的化感作用与资源竞争能力

为探讨反枝苋(Amaranthus retroflexus)在入侵农田生态系统过程中的化感作用与资源竞争能力之间的关系,采用添加活性炭的方法,比较了在有无活性炭条件下,反枝苋与大豆(Glycine max)的生物量积累、生物量分配及不同器官含氮量随供氮水平的变化情况。结果表明,在低供氮水平时,化感作用在竞争中占有的比重较大,在高供氮水平时,化感作用所占的比重相对降低;在高供氮条件下,反枝苋增加了对根和叶的生物量分配,而在低供氮条件下,则增加对繁殖器官的生物量分配;在氮素营养贫瘠的条件下,添加活性炭的大豆含氮量要明显小于不添加活性炭时,随着供氮水平的上升,差异逐渐减小。说明在入侵过程中,反枝苋可以通过调节自身的化感作用和生物量分配来适应不同的氮素营养条件。     

低磷营养胁迫下水稻化感抑草潜力的变化特性及其作用机理

在低磷(0.5 mg·L^-1) 营养胁迫下,运用生理生化方法分析了化感水稻PI312777（PI）与非化感水稻Lemont (Le) 对稗草抑制作用潜力的变化特性及其内在机理.结果表明,在低磷营养胁迫下,化感水稻品种PI对受体稗草根干重的抑制能力明显提高,在处理后的5 、10和15 d,其对稗草地下部干重的抑制率分别增加了5.64%、3.89%和12.13%,增加幅度比非化感水稻品种Le显著.生理生化分析结果表明,与正常营养条件相比,用低磷营养下生长的化感水稻PI的根系分泌物处理稗草5、10和15 d,受体稗草叶片中POD活性的促进率分别提高了20.19%、15.47%和6.68%,吲哚乙酸氧化酶活性的促进率分别提高了18.08%、17.71%和12.50%,硝酸还原酶活性的抑制率分别增加了13.89%、18.60%和2.10%. 在低磷营养胁迫下,化感水稻通过抑制受体植物的硝酸还原酶活性,影响其对氮营养的吸收,同时显著提高了吲哚乙酸氧化酶活性,减缓了受体稗草的生长速度,提高了其抑草作用潜力.     

不同水肥和光照条件对水稻化感特性的影响

采用盆栽实验对水稻化感品种华航1号在不同水肥和光照条件下的化感潜力和化感物质进行了研究。结果表明,华航1号在较高水肥条件下化感潜力较强,而在较低水肥条件下化感潜力下降,进一步对华航1号的化感特征物质的含量测定表明,在较低水肥条件下其化感物质没有显著的变化,只是次生物质的种类有所增加,尤其是一些具有抗病功能的次生物质的含量有所增加,在弱光照条件下华航1号的化感特征物质含量比在较强光照条件下要低,而且具有抗病功能的次生物质含量则保持较高水平,结果还发现,不论水、肥或光照条件的变化,华航1号对非伴生杂草的化感潜力都比伴生杂草要强。     

水稻化感生物测试方法的比较及应用

选用8个化感作用潜力各异的水稻品种(系)：PI312777、Lemont、Moroberekan、IAC25、IACA7、IAC120、Batatais、Iguape Cateto为供试材料,以田间伴生稗草为受体材料,比较分析了目前较为通用的3种生物测试方法：琼脂迟播共培法(RSA)、石英砂迟播共培法(RSS)和根系分泌物培养法(SRE)对水稻化感潜力的测试效果．结果表明,RSA法应用于测试水稻化感作用潜力的效果最好,RSS法和SRE法的测试结果虽较为一致,但两者都存在不足．据此,运用RSA法对57个水稻材料进行了化感抗草种质资源的初步筛选,从中获得5个强化感作用潜力的水稻材料：Iguape Cateto、PI312777、Azucena、Taichung Native 1和IAC25．     

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。     

红汁乳菇醇提物对水稻、稗草的化感影响

研究了红汁乳菇子实体乙醇抽提物对水稻、稗草的化感影响。用不同浓度的红汁乳菇子实体乙醇抽提物处理水稻和稗草种子 ,测量处理后不同时间的水稻和稗草幼苗的根长和苗高 ,按照 Willamson,G.B.的方法计算化感作用效应 (RI)。结果表明 :红汁乳菇乙醇抽提物对稗草幼苗的生长有显著的抑制作用 ,能够导致稗草叶片发黄 ,缩短稗草的根和苗 ,甚至延迟部分种子的萌发。其中 8.5 mg/ ml的乙醇抽提物处理稗草种子后 3d、8d,对根长的化感作用效应分别为 - 0 .98、- 0 .5 8;对苗高的化感作用效应分别为 - 0 .5 0、- 0 .5 1。 6 .5 m g/ ml和 10 .5 mg/ ml处理组的结果与 8.5 mg/ ml处理组的结果在 p=0 .0 5水平上差异不显著。对水稻幼苗的生长表现出一定的调节作用 ,在较低浓度下 (≤ 4 .5 0 mg/ m l)对水稻幼苗有一定促进作用 ,如 4 .5 0 m g/ ml处理水稻种子后 3d、8d,对苗高的化感作用效应分别为 0 .2 5、0 .10 ;在较高浓度下 (如≥ 6 .5 0 mg/ ml)对水稻根长和苗高均为抑制作用 ,6 .5 0 mg/ ml时抑制作用最强 ,处理水稻种子后 3d、8d对幼苗的根长的化感作用效应分别为 - 0 .93、- 0 .4 8;对苗高的化感作用效应分别为 - 0 .5 0、- 0 .2 2 ,在 p=0 .0 5水平上与其它处理组差异显著     

开放式空气CO2浓度增高条件下C3作物(水稻)与C4杂草(稗草)的竞争关系

通过田间试验,研究了FACE(开放式空气CO₂浓度升高)条件下C₃作物水稻(Oryza sativa)和C₄杂草稗草(Echinochloa crusgalli)的生长和竞争关系.结果表明,FACE条件下C₃植物水稻生物量和产量增加,叶片数增加,分蘖数增加,叶面积系数(LAI)增大;而C₄植物稗草相反.FACE条件下水稻和稗草叶面积均减少,而净同化率(NAR)均增加.FACE条件下水稻稗草比例为1:1时,水稻与稗草的生物量比率、产量比率、LAI比率、茎蘖比率和NAR比率均增加,水稻稗草的竞争关系发生变化,水稻(C₃植物)竞争能力增加,稗草(C₄植物)竞争能力下降.     

Genomic analysis of allelopathic response to low nitrogen and barnyardgrass competition in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

To explore the molecular mechanism of allelopathic rice in response to low nitrogen (N) supply or accompanying weed stress, allelopathic rice PI 312777 and its counterpart Lemont were grown under low N supply or co-cultured with barnyardgrass [Echinochloa crus-galli (L.) Beauv.] in hydroponics. The suppression subtractive hybridization (SSH) technique was employed to isolate the up-regulated genes in the treated rice accession. The results indicated that the expression of the genes associated with N utilization was significantly up-regulated in allelopathic rice PI 312777, and the higher efficiency of N uptake and its utilization were also detected in PI 312777 than that in Lemont when the two rice accessions were exposed to low N supply. This result suggested that the allelopathic rice had higher ability to adapt to low N stress than its non-allelopathic counterpart. However, a different response was observed when the allelopathic rice was exposed to accompanying weed (barnyardgrass) co-cultured in full Hoagland solution (normal N supply). It showed that the expression of the genes associated with allelochemical synthesis and its detoxification were all up-regulated in the allelopathic rice when co-cultured with the target weed under normal N supply. The results suggested that the allelopathic rice should be a better competitor in the rice-weed co-culture system, which could be attributed to increasing de novo biosynthesis and detoxification of allelochemicals in rice, consequently resulting in enhanced allelopathic effect on the target and preventing the autotoxicity in this process. These findings suggested that the accompanying weed, barnyardgrass is not only the stressful factor, but also one of the triggers in activating allelopathy in rice. This implies that the allelopathic rice is sensible of the existing target in chemical communication.     

Separation of allelopathy from resource competition using rice/barnyardgrass mixed-cultures

Plant-plant interference is the combined effect of allelopathy, resource competition, and many other factors. Separating allelopathy from resource competition is almost impossible in natural systems but it is important to evaluate the relative contribution of each of the two mechanisms on plant interference. Research on allelopathy in natural and cultivated plant communities has been hindered in the absence of a reliable method that can separate allelopathic effect from resource competition. In this paper, the interactions between allelopathic rice accession PI312777, non-allelopathic rice accession Lemont and barnyardgrass were explored respectively by using a target (rice)-neighbor (barnyardgrass) mixed-culture in hydroponic system. The relative competitive intensity (RCI), the relative neighbor effect (RNE) and the competitive ratio (CR) were used to quantify the intensity of competition between each of the two different potentially allelopathic rice accessions and barnyardgrass. Use of hydroponic culture system enabled us to exclude any uncontrolled factors that might operate in the soil and we were able to separate allelopathy from resource competition between each rice accession and barnyardgrass. The RCI and RNE values showed that the plant-plant interaction was positive (facilitation) for PI312777 but that was negative (competition) for Lemont and barnyardgrass in rice/barnyardgrass mixed-cultures. The CR values showed that one PI312777 plant was more competitive than 2 barnyardgrass plants. The allelopathic effects of PI312777 were much more intense than the resource competition in rice/barnyardgrass mixed cultures. The reverse was true for Lemont. These results demonstrate that the allelopathic effect of PI312777 was predominant in rice/barnyardgrass mixed-cultures. The most significant result of our study is the discovery of an experimental design, target-neighbor mixed-culture in combination with competition indices, can successfully separate allelopathic effects from competition.     

不同化感潜力水稻(Oryza sativa L.)苗期的氮素养分效率及相关基因的表达

为了阐明养分水平引起水稻(Oryza sativa L.)化感抑草潜力变化的生理生态机制,研究了不同N素营养处理下,不同化感潜力水稻苗期对N素营养逆境的响应特性及N素养分效率的差异,并运用实时荧光定量RT-PCR技术(FQ-PCR)检测与N素代谢和次生代谢关键酶的基因表达.结果表明:弱化感水稻品种Lemont对N素营养胁迫较敏感,强化感水稻品种PI312777对资源波动的适应性较强,N素养分效率较高.FQ-PCR分析结果显示,在低N条件下Lemont中的亚硝酸还原酶基因(nir),谷氨酰胺合成酶基因(gs)相对表达量均有不同幅度的下调, PI312777分别下调了1.2倍和1.4倍,而Lemont分别下调了3.0倍和1.8倍, Lemont下调的幅度分别是PI312777的2.5倍和1.3倍,但对于苯丙氨酸解氨酶基因(pal)与3-羟基-3甲基戊二酰辅酶A还原酶基因(hmgr)而言, PI312777叶组织中的pal和hmgr均上调表达,与对照相比上调了6.0倍和1.6倍,而Lemont中对应的基因均下调表达,分别下调了1.3倍和6.8倍,佐证了上述差异的分子生态学特性.     

Analysis of gene expressions associated with increased allelopathy in rice (Oryza sativa L.) induced by exogenous salicylic acid

The defense characteristics of allelopathic rice accession PI312777 and its counterpart Lemont induced by exogenous salicylic acid (SA) to suppress troublesome weed barnyardgrass (BYG) were investigated using the methods of suppression subtractive hybridization (SSH) and real-time fluorescence quantitative PCR (qRT-PCR). The results showed that exogenous SA could induce the allelopathic effect of rice on BYG and this inducible defense was SA dose-respondent and treatment time-dependent. PI312777 exhibited higher inhibitory effect than Lemont on BYG after treated with different concentrations of SA. The activities of cell protective enzymes including SOD, POD and CAT in the BYG plants co-cultured with PI312777 treated by SA were highly depressed compared with the control (co-cultured with rice without SA-treatment). Similar but lower depression on these enzymes except for CAT was also observed in the BYG plants when co-cultured with Lemont treated by SA. It is therefore suggested that allelopathic rice should be more sensitive than non-allelopathic rice to exogenous SA. Seventeen genes induced by SA were obtained by SSH analysis from PI312777. These genes encode receptor-kinase proteins, ubiquitin carrier proteins, proteins related to phenylpropanoid metabolism, antioxidant related proteins and some growth-mediating proteins. The differential expressions of these genes were validated in part by qRT-PCR in the two rice accessions. Our work elucidated that allelopathic rice possesses an active chemical defense and auto-detoxifying enzyme system such as the up-regulated enzymes involved in de novo biosynthesis of phenolic allelochemicals and the glutathione-S-transferase (GST) associated with xenobiotic detoxification.     

不同氮素供应下水稻酚类物质代谢关键酶基因差异表达

运用实时荧光定量PCR技术探讨了不同供氮条件下强化感与弱化感水稻苯丙烷代谢途径中9个关键酶基因的表达差异。结果表明,与正常氮素供应相比,低氮胁迫引起强化感水稻‘P1312777’中与酚类代谢途径相关的9个关键酶基因表达量均上调,表达量增幅在1.9～5.4倍之间,且以PAL基因上调倍数最大。而弱化感水稻‘Lemont’则相反,只有2个基因(苯丙氨酸裂解酶基因和肉桂酰CoA基因)表达上调,但上调倍数分别是强化感水稻对应的基因的22%和74%,其余的7个基因表达均下调,降幅在29%～72%之间,表明低氮胁迫诱发的水稻化感抑草能力增强与其体内酚类物质合成代谢增强有关。     

不同胁迫条件下化感与非化感水稻PAL多基因家族的差异表达

水稻苯丙氨酸解氨酶(PAL)调控酚酸类化感物质的合成代谢。编码PAL的基因是一个基因家族,包含至少11个基因成员,并受不同环境条件的调控。为了明确PAL基因家族中调控水稻化感作用的特定基因成员,本研究运用实时荧光定量PCR技术分析了低氮及稗草胁迫条件下强化感水稻PI312777与非化感水稻Lemont中根系的11个PAL成员基因的表达差异。结果表明,低氮和稗草胁迫条件下,PI312777和Lemont中的 PAL4和PAL10均不表达,其余9个PAL基因成员发生了不同程度的表达变化。其中,PAL11均上调表达,其分别在低氮处理和稗草胁迫的PI312777中上调3.29倍和1.07倍,而在相同处理下的Lemont中上调3.92倍和1.08倍;PAL3和PAL9则仅在低氮和稗草胁迫条件下的PI312777中上调表达,低氮胁迫分别为1.83倍和2.66倍,稗草胁迫为1.46倍和2.65倍;而这两个基因在相同处理下的Lemont中表达下调,低氮胁迫下调1.05和1.24倍,稗草胁迫下调1.14和1.16倍,推测PAL3和PAL9可能与胁迫初期调控水稻化感作用有关。     

外源水杨酸调控水稻化感抑草作用及其分子生理特性

为了探讨外源水杨酸（SA）调控水稻化感抑草效应的可行性,研究了不同浓度的外源SA对强化感水稻PI312777抑草效应的影响及其生理生化特性,并运用实时定量RT-PCR(FQ-PCR)技术检测SA介导的关键酶苯丙氨酸解氨酶基因（ZB8）的相对表达量.结果表明：外源SA能够诱导水稻化感抑草效应增强,而且这种诱导效应与SA的浓度和处理时间相关.叶面喷施SA后,PI312777对稗草的抑制率显著提高,其根系活力、超氧化物歧化酶（SOD）和过氧化物酶（POD）活性增强,过氧化氢酶（CAT）活性降低;而伴生杂草稗草的相应生理指标的变化趋势则相反.PI312777植株中的苯丙氨酸解氨酶（PAL）活性增强,总酚含量升高.0.2 mmol·L^-1的SA诱导水稻化感抑草效应最显著,该浓度下目的基因ZB8的相对表达丰度随处理时间先上调后下调,在24 h达到表达高峰.