中国境内稗(Echinochloa crus-galli)形态变异及其遗传和地理背景分析

采集了浙江、福建、江苏、湖南、湖北、四川、重庆、黑龙江、河南9个省的稗（Echinochloa crus-galli（L.）P.Beauv.）及其变种的33份种子,分别播种在相同的环境下,获得33个种群,测定了种群的16个形态性状,筛选出重复性好的9条ISSR引物,从33个种群中扩增出了109个位点。基于这些形态性状和ISSR位点信息,对33个种群先进行主成分分析,在此基础上再进行模糊均值聚类分析,探讨了它们的形态和遗传变化特点,及其与形态-遗传-地理背景三者之间的关系。主要结论如下：（1）33个种群可以鉴别出形态性状相对一致的4组,能够识别出西来稗（E.crus-galli var.zelayensis（Kunth）Farw.）、无芒稗（E.crus-galli var.mitis（Pursh）Peterm.）、细叶旱稗（E.crus-galli var.praticola Ohwi）;（2）基于109个位点信息对33个种群进行聚类分析得到了6组,部分组与形态聚类分组有一定的对应性;（3）33个稗草种群的遗传分化受地理背景因素的影响（r=0.684,n=33,P<0.001）;形态变异也有较明显的遗传背景因素（r=0.425,n=33,P<0.02）。在相对一致的稻田生境中,可能存在着形态上的趋同适应,使遗传上分化的组间在形态上又往往有交叉过渡,致使稗原变种（E.crus-galli var.crus-galli）、西来稗、无芒稗、短芒稗（E.crus-galli var.breviseta（Döll）Podp.）在形态上难以区别;（4）基于遗传和形态数据分析,发现细叶旱稗无论在形态上,还是遗传上,均形成了明显的一组,推测与该种长期适应于干旱生境有关,建议将细叶旱稗提升为种的水平,并将其命名为Echinochloa praticola（Ohwi）Guo S L,Lu Y L,Yin L P&Zou M Y。     

中国主要农区稻田稗草分类与多样性研究

将采自中国15个省和1个直辖市的206份稗属（Echinochloa）植物种子分别播种在相同栽培条件下,获得了206份子代样本及其21项形态性状数据,基于这些形态性状对206份稗属植物样本进行了聚类和主成分分析。结果表明：（1）从206份稗属植物样本中鉴别出了形态性状相对一致的8个分类群（G1-G8）,依次对应于湖南稗子（Echinochloa frumentaceae,2份样本）、水田稗（E.oryzoides,5份）、细叶旱稗（E.crus-galli var.praticola,6份）、硬稃稗（E.glabrescens,14份）、光头稗（E.colona,2份）、长芒稗（E.caudata,16份）、孔雀稗（E.cruspavonis,8份）、稗复合群（稗原变种E.crus-galli var.crus-galli、稗的变种无芒稗var.mitis、短芒稗var.breviseta和西来稗var.zelayensis,共150份）,还有3份样本没有聚合成组。它们依次占总样本量的0.97%、0.97%、2.42%、2.91%、3.88%、7.77%、6.80%、72.81%和1.46%,反映出中国主要农区稻田稗属植物主要以稗及其变种为主;（2）单因素方差分析表明,9个数量性状,包括叶长、叶宽、圆锥花序长度、总状花序长度、小穗芒长、小穗长度、第1颖长度/小穗长度、主茎直径和株高,在稗属植物的8个分类群（G1-G8）之间有显著差异;（3）根据8个分类群之间的形态性状特点及差异,给出了鉴别中国主要农区稻田稗属植物的分种检索表;（4）由于细叶旱稗具有总状花序简单、小穗无芒、叶较宽、植株高大、小穗紫黑色等独特形态性状,在206份稗属植物样本的聚类图上单独聚成一组,故建议将细叶旱稗作为独立种处理。     

尖角突脐孢菌与化学除草剂混用防治稻田稗草效果的研究

用平板表面萌发法测定化学除草剂对尖角突脐孢菌菌株X27分生孢子萌发的影响,在温室研究该菌分生孢子干粉与化学除草剂的相互作用,在田间评价其混剂的除稗效果.结果表明,二氯喹啉酸和苄嘧磺隆对分生孢子的萌发影响不明显,其它除草剂均有不同程度的影响.在温室条件下,菌X27干粉与二氯喹啉酸混用具有明显的增效作用,与敌稗混用具有加成作用;在水田条件下,单用菌X27干粉防除稗草效果差,防效只有60%,而与低量的二氯喹啉酸混用,防效明显提高,达90%以上.     

特异性适应稻作系统的稗草种群生活史特性

长期进行除草剂药效试验可能会导致田间杂草种群发生适应性进化。本研究在安徽南陵县除草剂药效试验专用稻田中采集了1个稗草种群A,并以从常规稻田采集的3个稗草种群为对照,开展同质园栽培试验。结果表明: 与3个对照种群相比,A种群稗草植株的单株种子产量显著减少,种子千粒重显著增加,幼苗生长速率显著加快,结实分蘖数显著增多,生育期显著缩短;A种群稗草成株的株高、生物量及对除草剂五氟磺草胺的敏感性均显著降低。A种群稗草幼苗3～4叶期时经五氟磺草胺推荐剂量2倍量(有效成分60 g·hm^-2)处理后,其株高、生物量及成熟种子产量(平均每株1066粒)显著降低,而抽穗期、结实分蘖数、单个总状花序的种子数及种子千粒重无显著差异。因此,种子较重、生活史周期短、植株矮小、结实分蘖多及对除草剂五氟磺草胺具有抗药性,使得A种群稗草对稻作系统具有特异适应性,应防止此类种群扩散至常规稻田。     

Competition between maize and Echinochloa crus-galli analysed by a hyperbolic regression model

Yield reduction of maize in relation to naturally established populations of Echinochloa crus-galli and Chenopodium album was studied in field experiments over 2 years in which the maize was grown at a wide range of weed densities. Both the crop and the weeds were harvested at intervals during the season. The competitive relations were described accurately by a model based on a hyperbolic relation between yield and plant density. The model can be linearised by considering the reciprocals of the average weight per plant. However, estimating the regression coefficients by linear regression introduced a severe bias due to heterogeneity of variances. Estimation was improved by applying non-linear regression, using a logarithmic transform of the yield equation. Fitted regressions were used to interpolate the yield data to standard weed densities. At a density of 100 Echinochloa plants m-², maize yields were reduced by 8% and 82% in 1982 and 1983, respectively, illustrating the problems in generalising the results of one competition experiment to the other. Three possible fields of application of the competition model are discussed, i.e. adjustment of experimental plot yield for variation in weed population, prediction of expected crop yield losses, and prediction of long-term changes in weed seed populations.     

Bioactive steroids from Oryza sativa L

Rice is one of the most interesting crops in the world from both the social and the economic point of views. The monoculture practices along with the heavy use of herbicides are characteristic of modern agriculture and are inducing the appearance of tolerant and/or herbicide resistant weed biotypes. This is the case the world's main weed of rice barnyardgrass (Echinochloa crus-galli). Alternative strategies for weed suppression consist of the use of chemicals from rice due to necessity of obtaining new herbicides with new modes of action that could prevent resistance phenomena. In order to carry out a study that guides to the isolation of the most active compounds from rice, different extracts were achieved, and their activities evaluated. So, all the plant material was divided into three parts: fresh plant, dried plant, and fresh plant from Pluviotron. The aerial part was separated from roots in all cases and extracted in water, in organic solvents as well as with the Pluviotron device. The activity of the 12 extracts obtained was evaluated using a generalist bioassay, wheat etiolated coleoptiles bioassay, and a phytotoxic bioassay on barnyardgrass as target species. The bioactive extracts were fractionated and 15 compounds were isolated and identified by spectroscopic methods. Eight of these compounds were isolated for the first time in Oryza sativa. The most phytotoxic compounds on E. crus-galli were ergosterol peroxide and 7-oxo-stigmasterol. In the case of ergosterol peroxide the activity was higher than the commercial herbicide Logran. This is the first report of potential allelopathic activity of steroids on weeds based on their phytotoxicity.     

Effect of increased concentration of soil CO2 on intermittent flushes of seed germination in Echinochloa crus-galli var. crus-galli

Soil-buried seeds of barnyardgrass ( Echinochloa crus-galli var. crus-galli ) germinated from April to June in three intermittent flushes. The later two flushes of germination occurred after heavy rainfall. Carbon dioxide concentration in soil air transiently increased to 30 dm³ m^–3 after the rainfall, probably due to the increase in soil temperature and water potential. Germination of exhumed seeds was stimulated by exposure to CO₂ at 30 dm³ m^–3. Fluctuating temperature, light, water, ethylene, and nitrate are known to promote seed germination in many species. However, of these environmental factors, within ranges found in the field, only CO₂ was effective in enhancing the germination of barnyardgrass seeds. We conclude that soil CO₂ is responsible for causing intermittent flushes of germination. Detection of vegetation gaps may be explained by the responsiveness of buried seeds to CO₂.     

不同水层和密度条件下化感作用水稻对无芒稗的干扰控制作用

不同水层下水稻对无芒稗的干扰控制作用研究表明,水稻化感作用品种吓一跳、谷梅2号和中156对无芒稗的株高抑制作用比无化感作用品种秀水63和春江11呈显著性差异．随盆栽水稻密度增加,水稻对无芒稗的干扰控制作用加大．在每盆4株栽植密度下,水稻化感作用品种鸡早籼、谷梅2号显著抑制无芒稗植株株高;在每盆32株密度时,水稻化感作用品种吓一跳、PI312777、TN1等对受控无芒稗的干扰控制与无化感作用品种春江11相比,呈显著性水平．     

Constitutive and Inducible Aerobic and Anaerobic Stress Proteins in the Echinochloa Complex and Rice

Anaerobic stress resulted in a change in the protein accumulation patterns in shoots of several Echinochloa (barnyard grass) species and Oryza sativa (L.) (rice) as resolved by two-dimensional gel electrophoresis. Of the six Echinochloa species investigated, E. phyllopogon (Stev.) Koss, E. muricata (Beauv.) Fern, E. oryzoides (Ard.) Fritsch Clayton, and E. crus-galli (L.) Beauv. are tolerant of anaerobiosis and germinate in the absence of oxygen, as does rice. In contrast, E. crus-pavonis (H.B.K.) Schult and E. colonum (L.) Link are intolerant and do not germinate without oxygen. Computer analysis of the protein patterns from the four tolerant species and rice indicated that the anaerobic response is of five classes: class 1 proteins, enhanced under anaerobiosis (9 to 13 polypeptides ranging from 16-68 kD); class 2 proteins, unique to anaerobiosis (1 to 5 polypeptides ranging from 17-69 kD); class 3 proteins, remained constant under aerobiosis and anaerobiosis; class 4 proteins, prominent only in air and repressed under anoxia (3 to 7 polypeptides ranging from 19-45 kD); and class 5 proteins, unique to aerobiosis (1 to 4 polypeptides ranging from 18-63 kD). In the intolerant species, E. colonum and E. crus-pavonis, no polypeptides were enhanced or repressed under anoxia (class 1 and class 4, respectively), whereas in the tolerant Echinochloa species and rice, a total of at least 9 to 13 anaerobic stress proteins and 4 to 7 "aerobic" proteins were noted. Immunoblotting identified two of the major anaerobic stress proteins as fructose-1,6-bisphosphate aldolase and pyruvate decarboxylase. Based on the differential response of the intolerant species to anaerobiosis, we suggest that another set of genes, whose products may not necessarily be among the major anaerobic stress polypeptides, might confer tolerance in Echinochloa under prolonged anaerobic stress.     

Inhibition of Barnyardgrass 4-Hydroxyphenylpyruvate Dioxygenase by Sulcotrione

4-Hydroxyphenylpyruvate dioxygenase (EC 1.13.11.27) was partially purified from barnyardgrass (Echinochloa crus-galli L.) leaves and assayed by high-performance liquid chromatography analysis of product formation or by the capture of released 14CO2. The bleaching herbicide sulcotrione [2-(2-chloro-4-methanesulfonylbenzoyl)-1,3-cyclohexanedione] was shown to be a potent, linear competitive inhibitor of 4-hydroxyphenylpyruvate dioxygenase. Kinetic analyses determined that the Km for the substrate, 4-hydroxyphenylpyruvate, was 4.3 [mu]M, and the Ki value was 9.8 nM for sulcotrione.     

二氯喹啉酸诱导稗草EcDnaJ基因表达

DnaJ作为分子伴侣在植物抗逆中起重要作用. 但目前在二氯喹啉酸逆境下,其在抗药性稗草中表达特点却鲜有报道. 本研究采用RACE技术从抗二氯喹啉酸稗草中克隆了1个DnaJ基因, 命名为EcDnaJ1 (GenBank登录号：JX518598), 其cDNA全长为2 154 bp, 开放阅读框为1 350 bp,编码449 个氨基酸, 理论分子量为48.4 kD, 等电点为9.5. 该蛋白质氮端含有1个保守的J结构域, 中部含有4个模式为CxxCxGxG的锌指结构. Real-time PCR分别测定EcDnaJ1在二氯喹啉酸抗性和敏感的稗草生物型苗期叶、根及成株期根、茎、叶和种子中的表达, 在感、抗生物型的相对表达量分别是0.8～20.9和7.4～30.2, 其中在抗性稗草苗期叶片中相对表达量最高为30.2, 而在敏感稗草种子中最低为0.8, 抗性稗草是敏感稗草的1.4～9.2 倍. 受二氯喹啉酸诱导后, 其在感、抗稗草的相对表达量分别为35.8～72.5和84.9～261.9, 在抗性稗草苗期叶片中相对表达量最高为261.9, 而在敏感稗草的种子中相对表达量最低为35.8, 抗性稗草是敏感稗草的2.4～3.6 倍. 诱导前后, 无论是在苗期根和叶中还是成株期的根、茎、叶和种子中, EcDnaJ1表达量均是抗性稗草高于敏感稗草. 抗、感稗草生物型的EcDnaJ1在mRNA水平差异表达暗示,它可能参与了稗草对二氯喹啉酸的抗药性.     

氮素增加条件下丛枝菌根真菌对陆稻和无芒稗相互作用的调节

采用盆栽方法研究了增氮条件下丛枝菌根真菌(AMF)对无芒稗(Echinochloa crusgallivar.mitis L.)和陆稻(Oryza sativa L.)相互作用的调节.结果表明:在单种条件下,无芒稗的菌根侵染率增加,陆稻的菌根侵染率降低;无AMF和接种AMF处理陆稻的生物量分别提高13.48%和42.35%,总磷吸收分别提高2.55%和4.07%,总氮吸收分别提高62.09%和30.35%;无芒稗的生物量分别提高15.65%和20.24%,总磷吸收分别提高4.06%和3.88%,总氮吸收分别提高30.35%和15.10%.在混种条件下,无芒稗的菌根侵染率增加,而陆稻无显著变化;无AMF和接种AMF的无芒稗与陆稻的生物量比值降低,总氮吸收比值无显著变化,无AMF处理的总磷吸收比值增加,而接种AMF处理降低.表明增氮条件下AMF提高了无芒稗对陆稻的竞争.     

应用主成分分析法和最小生成树法研究浙中秋旱作物田杂草种间生态关系

研究意义浙江金华地处浙江中部,旱地面积有５．０９×１０４ｈａ,占耕地总面积的１８％以上,棉花、大豆、甘薯等秋旱作物是该地区重要的经济作物,几年来,由于农村劳动力转移,中耕除草减少,草害问题日趋严重。化学除草是解决旱田草害的有效方法。但是,除草剂的长期...     

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.     

Photosynthesis and physiology responses of paired near-isogenic lines in waxy maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) to nicosulfuron

Nicosulfuron is a post-emergence herbicide used for weed control in fields of maize (Zea mays L.). We used a pair of nearly isogenic inbred lines, SN509-R (nicosulfuron-resistant) and SN509-S (nicosulfuron-sensitive), to study the effect of nicosulfuron on waxy maize seedling. After the nicosulfuron treatment, net photosynthetic rate, stomatal conductance, transpiration rate, leaf maximum photochemical efficiency of PSII, photochemical quenching of chlorophyll fluorescence, and the actual photochemical efficiency of PSII were significantly lower in SN509-S than those of SN509-R, contrary to intercellular CO₂ concentration, stomatal limitation, and nonphotochemical quenching. Compared to SN509-R, antioxidant enzyme activities in SN509-S decreased significantly in response to the nicosulfuron treatment, while SN509-S exhibited an increased malondialdehyde content, which was associated with lower antioxidant enzyme activities. These results collectively suggest that the nicosulfuron-resistance mechanism was associated with photosynthetic rate, reactive oxygen species metabolism, and protective mechanisms.     

Comparison Study of Allelochemicals and Bispyribac-Sodium on the Germination and Growth Response of Echinochloa crus-galli L.

The phytotoxic effects of two allelochemicals (trans-cinnamic acid and syringaldehyde) at different concentrations (1000, 100, 10, and 1 µM) on seed germination, seedling growth, and physiological and biochemical changes of Echinochloa crus-galli L. were tested by comparison to a commercial herbicide ‘Nominee’ (that is, 100 g/L bispyribac-sodium). trans-Cinnamic acid and the herbicide inhibited seed germination completely at 100 µM, whereas for syringaldehyde, complete inhibition required 1000 µM. However, with 100 µM syringaldehyde, the seed germination of the test species was 53% of the control. Allelochemicals and the herbicide delayed seed germination and significantly affected the speed of germination index (S), speed of cumulative germination index (AS), and coefficient of germination rate (CRG). The roots were more affected when nutrients were not added to the growth bioassay. In general, with the increasing concentration of allelochemicals from 100 to 1000 µM, the inhibitory effects increased. Via microscopy analysis, we found leaf blade wilting and necrosis at concentrations above 100 µM in allelochemical-treated plants. Roots of E. crus-galli treated with 1000 µM allelochemicals had black points on root nodes but had no root hairs. The anatomy of roots treated with allelochemicals (1000 µM) showed contraction or reduction of root pith cells as well as fewer and larger vacuoles compared to the control. The allelochemicals also showed remarkable effects on seedling growth, SPAD index, chlorophyll content, and free proline content in a pot culture bioassay, indicating that trans-cinnamic acid and syringaldehyde are potent inhibitors of E. crus-galli growth and can be developed as herbicides for future weed management strategies.

     

Effects of nicosulfuron on the abundance and diversity of arbuscular mycorrhizal fungi used as indicators of pesticide soil microbial toxicity

The key role of arbuscular mycorrhizal (AM) fungi in maintaining soil fertility and ecosystem functioning and their general sensitivity to pesticides make them good candidate bioindicators in pesticide soil microbial toxicity assessment. We investigated the impact of the herbicide nicosulfuron on mycorrhizal colonization and community structure of AM fungi via a pot-to-field experimental approach. This allowed the assessment of nicosulfuron toxicity (i) at extreme exposure schemes (pot experiment, Tier I) invoked by the repeated application of a range of dose rates (x0, x10, x100, x1000 the recommended dose) and (ii) under realistic exposure scenarios (x0, x1, x2, x5 the recommended dose) in the field (Tier II). In the pot experiment, the x100 and x1000 dose rates significantly reduced plant biomass, mycorrhizal colonization and AM fungal richness as determined by DGGE. This coincided with the progressive accumulation of herbicide concentrations in soil. In contrast, no effects on AM fungi were observed at the nicosulfuron dose rates tested in the field. Clone libraries showed that the majority of AM fungi belonged to the Glomus group and were sensitive to the high levels of nicosulfuron accumulated in soil at the latter culture cycles. In contrast, a Paraglomeraceae and a Glomus etunicatum ribotype were present in maize roots in all cycles and dose rates implying a tolerance to nicosulfuron-induced stress. Overall, the deleterious effects of nicosulfuron on AM fungi induced by the highest dose rates in the pot experiment could be attributed either to fungal-driven toxicity or to plant-driven effects which have subsequent implications for mycorrhizal symbiosis. We suggest that the tiered pot-to-field experimental approach followed in our study combined with classic and standardized molecular tools could provide a realistic assessment of the toxicity of pesticides onto AM fungi as potential bioindicators.     

稗属杂草对水稻生长发育和产量的影响

以新两优6号(籼稻)和南粳46(粳稻)为材料,自水稻移栽至成熟分别与无芒稗(T₁)、稗(T₂)、西来稗(T₃)和光头稗(T₄)共生,稗草密度为6株·m^-2,以无稗草水稻处理为对照,研究不同稗草对水稻生长发育和产量的影响.结果表明：与对照相比,不同稗草对水稻的干扰表现不同,T₁、T₂、T₃处理使籼稻产量分别下降19.2%、10.8%、21.9%,使粳稻产量分别下降39.7%、25.3%、47.3%,但T₄处理对2个水稻品种的产量均无显著影响.水稻和稗草共生过程中,T₁、T₂和T₃处理显著降低了水稻成熟期的干物质积累量及灌浆期的剑叶光合速率、根系氧化力和籽粒ATP酶活性,降低幅度为T₃>T₁>T₂,T₄处理与对照差异不显著;各处理对水稻最终分蘖数和株高均无显著影响.表明4种稗草对水稻生长的影响由强到弱表现为：T₃>T₁>T₂>T₄,稻 稗共生时水稻剑叶光合速率、根系氧化力和籽粒ATP酶活性降低是导致水稻生产力下降的重要原因.     

Enantioselective effects of optically active alpha-methylbenzyl-s-triazine on the root growth of rice and Echinochloa plants and their herbicidal activity

The chiral requirement on the alpha-methylbenzyl moiety of 2,4-diamino-6-chloro-s-triazine for sufficient inhibition of root growth was similar towards both rice and barnyard millet. With the monoalkylamino series, the most suitable configuration was markedly changed by the substituent on the other amino moiety. However, for the dialkylamino series, the (S)-enantiomer was an active inhibitor. Clear species selectivity between rice and barnyard millet was observed in the series for the (R)-enantiomers, providing high herbicidal activity toward Echinochloa plants and safety toward rice. The enantioselectivity against barnyard millet increased with increasing inhibitory activity of the active enantiomers, following Pfeiffer's rule. R-EtNH (3) controlled the growth of barnyardgrass with leaf-burning (LB) under paddy conditions, and S-EtNH (4) and S-Et2N (20) controlled the growth without LB. The RS-EtNH derivative is an interesting inhibitor controlling the growth of barnyardgrass from the just-germinated stage (by the (R)-enantiomer) to early-middle growth stage (by the (S)-enantiomer).