干旱胁迫下内生真菌感染对黑麦草光合色素和光合产物的影响

以含有内生真菌的黑麦草 (L olium perenne L.)种子为材料 ,采用加热处理方式构建内生真菌非感染的黑麦草种群 ,通过比较内生真菌感染 (EI)和非感染 (EF)植株在正常条件下和干旱胁迫条件下叶片相对水分含量、叶绿素、可溶性糖和淀粉含量等指标的差异 ,探讨黑麦草 EI和 EF种群对干旱胁迫的适应性差异。结果表明 :在中度胁迫后期 ,EI植株叶片的 RWC显著高于 EF植株 ,即 EI植株的保水能力更强。轻度水分胁迫下 ,内生真菌感染可使其宿主植物的可溶性糖含量增加 ,以增强宿主的渗透调节能力 ,随着干旱胁迫强度的加大 ,内生真菌的这一增益效应不再起作用 ,此时 ,宿主植物将更多的光合产物——淀粉积累于体内 ,以度过不良环境。第 2年春天 EI和 EF种群的恢复生长情况进一步表明 ,经过中度干旱胁迫后 ,EI种群的恢复更为迅速。生物量的大小是植物种群净光合作用能力的直接体现 ,研究中在中度干旱胁迫条件下 ,黑麦草 EI种群的生物量显著高于EF种群 ,但从光合色素的变化来看 ,相同水分状况下 EI和 EF植株的 Chla、Chlb以及 Car的变化趋势比较接近 ,这说明内生真菌感染并未缓解干旱胁迫对光合色素的破坏 ,内生真菌可能通过其它途径来改善宿主植物的光合能力     

Photosynthetic pigments and photosynthetic products of endophyte-infected and endophyte-free Lolium perenne L. under drought stress conditions

Endophyte-infected (EI) seeds of Lolium perenne L. were used to attain endophyte-free (EF) population by heating the seeds at 43°C for 15 min and then 57°C for 25 min. Relative water content (RWC), chlorophyll, soluble sugar and starch content of EI and EF populations under normal and drought stress conditions were compared to investigate the effect of endophyte infection on the host plant. Under severe stress, RWC of EI leaf was significantly higher than that of EF leaf, i.e. EI plants took more advantages over EF plants in water-holding ability. Under mild stress, endophyte could enhance soluble sugars in host plants to improve their osmotic ability. With stress intensification, the improvement of endophyte no longer existed, and more photosynthetic products (such as starch) accumulated in EI plants to survive through the undesirable conditions. In the next spring, EI populations will recover more rapidly than EF populations. The biomass of a population is closely related to its photosynthesis. Under severe stress, EI population significantly accumulated more biomass than EF population. As far as photosynthetic pigments were concerned, contents of Chla, Chlb and Car of EI plants were close to those of EF plants, which suggested that endophyte infection didn’t alleviate photosynthetic pigments from being destroyed by drought stress, and endophyte might improve photosynthesis ability of its host plant in other ways. __________ Translated from Acta Ecologica Sinica, 2005, 25 (2) [译自: 生态学报, 2005, 25(2)]     

Photosynthetic pigments and photosynthetic products of endophyte-infected and endophyte-free Lolium perenne L. under drought stress conditions

Endophyte-infected (EI) seeds of Lolium perenne L. were used to attain endophyte-free (EF) population by heating the seeds at 43°C for 15 min and then 57°C for 25 min. Relative water content (RWC), chlorophyll, soluble sugar and starch content of EI and EF populations under normal and drought stress conditions were compared to investigate the effect of endophyte infection on the host plant. Under severe stress, RWC of EI leaf was significantly higher than that of EF leaf, i.e. EI plants took more advantages over EF plants in water-holding ability. Under mild stress, endophyte could enhance soluble sugars in host plants to improve their osmotic ability. With stress intensification, the improvement of endophyte no longer existed, and more photosynthetic products (such as starch) accumulated in EI plants to survive through the undesirable conditions. In the next spring, EI populations will recover more rapidly than EF populations. The biomass of a population is closely related to its photosynthesis. Under severe stress, EI population significantly accumulated more biomass than EF population. As far as photosynthetic pigments were concerned, contents of Chla, Chlb and Car of EI plants were close to those of EF plants, which suggested that endophyte infection didn’t alleviate photosynthetic pigments from being destroyed by drought stress, and endophyte might improve photosynthesis ability of its host plant in other ways.     

渗透胁迫下内生真菌感染对黑麦草幼苗生长的影响

以含有内生真菌的黑麦草(Lolium perenne L．)种子为材料,采用4C冰箱内和20℃光照培养箱内保存18个月的方式分别构建内生真菌感染(EI)和内生真菌非感染(EF)的黑麦草种群,通过比较EI和EF种群在正常条件下(对照)和渗透胁迫条件下种子发芽、幼苗生长等方面的差异,探讨内生真菌对其宿主植物的直接和间接影响。结果表明：在对照和胁迫条件下,E1种子的发芽势均明显高于EF种子,而只在重度渗透胁迫下,E1种子的发芽率才显著高于EF种子。对于黑麦草幼苗而言,渗透胁迫下内生真菌对宿主植物的地上部分和地下部分均有增益作用,最终表现为E1种群的总生物量显著高于EF种群,其中对地上部分的促进作用表现为内生真菌的存在不仅提高了宿主叶片的延伸速率、使EI叶片比EF叶片更长、叶面积更大,而且在重度胁迫下,E1种群的分蘖数也显著高于EF种群;对地下部分的促进作用表现为EI种群的根系总长度和根干重均高于EF种群。     

干旱胁迫下内生真菌感染对黑麦草叶内几种同工酶的影响

以内生真菌感染(endophyte-infected,EI)与不感染(endophyte-free,EF)的黑麦草(Lolium perenne L．)种子建立实验种群,分别对其施加长时间不同强度的干旱胁迫,通过比较黑麦草体内过氧化物酶(POD)、超氧化物歧化酶(SOD)、多酚氧化酶(PPO)活性及其同工酶谱的变化以探讨保护酶系统在内生真菌——植物共生体的抗旱性方面所作的贡献。研究结果表明,水分胁迫和内生真菌对黑麦草3种酶的影响不仅表现在总量上而且表现在同工酶的酶谱及各区带的酶活力上。就总酶活力而言,EI和EF植株中POD、SOD和PPO的活性均随着干旱胁迫强度的增加而增加,进一步将EI和EF植株的酶活力进行比较,发现与EF植株相比,EI植株中POD和PPO的活性相对较低,而SOD的活性相对较高。从同工酶的谱带数量和强弱来看,POD同工酶各区带活力均随干旱胁迫强度的增加而增加,EI植株叶片增加的幅度高于EF叶片,而且EI叶片在重度胁迫下出现了1条新带SOD同工酶各区带活力在EI叶片中有随干旱胁迫增加而增加的趋势,而在EF叶片中有些区带酶活力增强,有些区带酶活力减弱,且EI叶片在中度胁迫下出现了1条新带;PPO同工酶随干旱胁迫的增强,EI和EF叶片均表现为有些区带酶活力增强,有些区带酶活力减弱。总之,内生真菌的感染虽然没有显著提高宿主植物黑麦草POD、SOD和PPO的活性,但使宿主黑麦草对干旱胁迫的反应更为迅速,其中既包括POD、SOD等酶活力的迅速升高,也包括新酶带的产生。     

土壤干旱条件下保水剂对多年生黑麦草光合特性的影响

以多年生黑麦草（Lolium perenne Linn．）为研究对象,对土壤干旱条件下（土壤含水量12．6％、10．5％、8．4％和6．3％）使用保水剂后叶片光合特性的动态变化进行了分析。结果显示：随土壤含水量的降低和胁迫时间（12d）的延长,多年生黑麦草叶片的净光合速率（Pn）、气孔导度（Gs）、蒸腾速率（Tr）和光能利用效率（LUE）呈逐渐下降的趋势;随土壤含水量的降低,胞间CO2浓度（Ci）逐渐升高、瞬时水分利用效率（WUE）逐渐降低,但随胁迫时间的延长Ci和WUE则呈现不同的变化趋势;总体上,在土壤含水量不同的条件下及不同的胁迫时间各指标均有显著差异（P〈0．05）。与未添加保水剂的各处理组相比,添加质量分数2％保水剂后多年生黑麦草叶片的Pn、Gs、WUE和LUE值均增大,而Ci和Tr值则降低,差异达显著水平（P〈0．05）。研究结果表明：合理使用保水剂能提高多年生黑麦草叶片的光合能力以及土壤的保水能力,增强多年生黑麦草对干旱环境的适应性。     

水分胁迫下内生真菌感染对黑麦草叶内游离脯氨酸和脱落酸含量的影响

以黑麦草为实验对象,研究了干旱胁迫条件下内生真菌感染对植株叶片含水量和叶内游离脯氨酸含量的影响,同时对渗透胁迫条件下植株叶内ABA含量的变化进行了分析。结果表明：①内生真菌的感染有助于使叶片保持较高的含水量;②在两种形式的水分胁迫下,。前期至中期高感染种群的叶片游离脯氨酸含量低于感染种群,而在末期则有高出低感染种群的趋势;③内生真菌感染对黑麦草叶内ABA累积的正效应只发生在轻度渗透胁迫下的较短时间范围内。     

重金属递进胁迫对黑麦草初期生长的影响

通过研究Cu²⁺、Zn²⁺、Cd²⁺与Pb²⁺胁迫对黑麦草初期生长的影响,结果表明：4种重金属对种子发芽率抑制效应相对较小,尤其Cu²⁺与Zn²⁺的抑制作用最小。高浓度Cu²⁺、Cd²⁺胁迫对株高、根系长度、地上生物量的抑制作用相对较大,尤其Cu²⁺对根系生长的抑制效应最大,在300 mg·L^-1下,与对照相比,根长最高下降了 85.48%。高浓度Cd²⁺胁迫显著降低了叶绿素含量,在300 mg·L^-1时比对照降低了45.51%;与对照相比,Cu²⁺与Zn²⁺所有处理都增加了叶绿素含量。从递进胁迫进程看,一些重金属对某一生长指标的影响往往表现在低浓度具有促进作用,而高浓度又存在明显的抑制效应。     

黑麦草内生真菌感染状况的检测及定量分析

选取内生真菌特异性引物,成功建立了利用PCR技术对黑麦草中内生真菌感染状况的检测和定量分析方法。此检测方法的准确性高于常规乳酸-苯胺蓝染色法。利用实时荧光PCR定量分析的结果表明：不同植株之间内生真菌含量差异较大,而同株植物相同龄级分蘖之间内生真菌含量无显著差异。由此可见内生真菌的含量不仅与植物种以及品种有关,也与植物的基因型密切相关。     

Partitioning of sugars in Lolium perenne (perennial ryegrass) during drought and on rewatering

Simulated swards of perennial ryegrass ( Lolium perenne ) growing in 1-m³ soil blocks in the glasshouse were either well watered or deprived of water for 57 d and then rewatered. The first aim was to measure effects of drought on sugar (water-soluble carbohydrate) composition of laminae and sheaths of mature laminae, and bases and laminae of young (growing) leaves. The second aim was to use pulse labelling with ¹⁴CO₂ to follow the partitioning of recently-fixed assimilates, and the assembly and consumption of reserve sugars (fructans). Over the last 7 d of drought growth almost stopped, old leaves died faster than they were replaced, and total sugar (which had doubled in concentration during drought) was rapidly consumed. Leaf laminae had lower content of total sugars and of large fructan (DP>5) than did growing bases and sheaths. Drought greatly reduced the rate at which sugar was exported from the laminae to the sheaths and growing leaf bases, and the rate at which it was converted to fructan. Nevertheless, fructan accumulated over the first 50 d of drought. Rewatering did not result in depolymerization and remobilization of sugars that had been formed during the last 7 d of drought, but stimulated their further assembly into high-DP fructans. Our hypothesis, that accumulation of neo-kestose (a DP-3 fructan) in droughted laminae was a symptom of sugar remobilization just before death, was disproved. It is concluded that sugar reserves contribute to drought resistance only under extreme conditions. The specific role of fructan in dry environments might be to improve regrowth when drought is relieved, rather than to enhance growth during drought.     

Field performance of cell-suspension-derived Lolium perenne L. regenerants and their progenies

 Two sets of plants (Lb and Lc), regenerated from different single-genotype-derived embryogenic suspension cultures of Lolium perenne cv Citadel, were evaluated for agronomic traits in a modified polycross design in the field. Seed from the primary regenerated plants was harvested to evaluate morphological and phenological traits of corresponding progenies in a replicated field experiment. When compared to seed-grown plants of the same cultivar, primary regenerants of the Lb set showed a significant delay in ear emergence and a more-erect growth habit, while primary regenerants from the Lc set showed a significantly higher seed yield. However, progenies of regenerated plants did not differ from those of seed-grown plants. Embryogenic suspension cells of L. perenne have the potential for producing fertile, well-performing, material which can be integrated into breeding programs. Received: 3 November 1997 / Accepted: 25 November 1997     

黑麦草(Lolium perenne L.)内生真菌的检测、分离及鉴定

从多年生黑麦草(LoliumperenneL.)5个品种———SR4000、Pinnacle、Topgun、CalypsoⅡ、Justus中分离出61个菌株。次培养后,所得形态稳定的菌株可分为4个形态群,依据其形态特征及APPCR的结果,确定其中的57个分离菌株为Neotyphodiumlolii。     

外源-氧化氮供体硝普钠对黑麦草幼苗耐碱性的影响

采用营养液砂培方法,研究外源一氧化氮(NO)供体硝普钠(SNP)对NaHCO3胁迫下黑麦草幼苗生长、活性氧代谢和渗透溶质积累的影响.结果表明:60 μmol·L-1 SNP能够缓解100 mmol·L-1NaHCO3胁迫对黑麦草幼苗生长的抑制作用,减缓NaHCO3胁迫导致的叶片O2产生速率、H2O2和丙二醛(MDA)含量的增加,提高NaHCO3胁迫下幼苗叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)、抗坏血酸过氧化物酶(APX)、质膜H+-ATP酶的活性和谷胱甘肽(GSH)、可溶性糖、可溶性蛋白质和脯氨酸的含量及K+/Na+比,降低过氧化氢酶(CAT)活性和抗坏血酸(AsA)含量,对游离氨基酸含量影响不大.上述结果表明,NO可能通过激活抗氧化系统活性、促进渗透溶质积累和改善Na+、K+平衡等缓解碱胁迫对幼苗的伤害,从而提高黑麦草的耐碱性.     

黄土性土壤固化对黑麦草生长和根系活力的影响

采用盆栽试验,研究了安塞黄绵土不同容重、不同固化剂(路邦EN-1固化剂)掺量对黑麦草生长和根系活力的影响.结果表明:黄绵土容重在1.2～1.4 9·cm-3范围内,随土壤容重的增加,黑麦草叶绿素含量、根系活力、根冠比、根生物量和植株生物量均降低;各土壤容重条件下,黑麦草叶绿素含量、根系活力、根冠比、根生物量和植株生物量均高于对照,且随着固化剂掺量的增加均呈先增加后降低的趋势.土壤容重和固化剂掺量交互作用对根生物量和总生物量的影响均显著(P＜0.05).总体来看,土壤容重1.3 g·cm-3、固化剂掺量0.15％处理下,各指标值均最高.     

不同氮素营养条件下内生真菌感染对黑麦草光合和蒸腾速率及生物量的影响

以黑麦草(Lolium perenne L)为实验材料,研究在不同氮素营养条件下内生真菌(Neotyphodium lolii)感染对其光合特性及生物量的影响.结果表明:1)整个实验中,内生真菌感染对黑麦草净光合速率、蒸腾速率以及水分利用效率均无显著影响,但在中氮和高氮水平时,感染种群的净同化累积值有超过非感染种群的趋势;2)在本实验的第一处理期和第二处理期,内生真菌感染对黑麦草绿色部分的干物质产量无显著影响,但在第三处理期感染种群的绿色部分的干物质产量显著高于非感染种群.     

盐胁迫下两草种SOD和POD及脯氨酸动态研究

在NaCl、KCl、MgSO₄及其复合盐胁迫下, 研究草坪草-金牌美达丽(Lolium perenne)和猎狗(Fesluca elata) 中脯氨酸含量、超氧化物歧化酶和过氧化物酶活性的变化。结果表明:不同盐分、不同浓度胁迫下脯氨酸含量呈波动式变化, 其峰值出现在较高浓度下, 说明在一定范围内盐胁迫强度越大脯氨酸积累越明显;SOD 的最高值出现在低浓度下, 而最低值无此特点, 不同盐胁迫下的最高值、最低值相差不多, 说明L.perenne 和F.elata 相差不明显;POD 活性随盐浓度增加呈波动性变化, L.perenne (除KCl)和F.elata (除MgSO₄)最高值均出现在低浓度下, L.perenne 的最低值集中在高浓度, 而F.elata 的最低值则集中在低浓度, F.elata在高浓度胁迫下, 处于一个比较稳定的水平, 高于或接近CK, 且各值比较接近, 而在L.perenne 中却没有这一结果。     

Neighbourhood effect of genotypes of Rhizobium leguminosarum biovar trifolii, Trifolium repens and Lolium perenne

 Ryegrass, white clover and Rhizobium isolated from the corresponding clover nodules, were harvested from a natural pasture in the Massif Central mountains (France). The specificity between Lolium, Trifolium and Rhizobium, and the genetic diversity of Rhizobium were examined. This study showed that: 1) Natural neighbouring combinations of white clover and ryegrass, re-planted together in pots, accumulated a higher biomass than non-neighbouring ones. This increase of mass is higher in the presence of the native strain of Rhizobium. 2) When white clover was inoculated with a mixture of Rhizobium strains, nodules were more often formed by its native strain. 3) The genetical diversity of the Rhizobium leguminosarum biovar trifolii was very high, as revealed by electrophoresis of esterases on seven substrates. These results support the hypothesis that there is a co-adaptation between white clover, ryegrass and Rhizobium Received: 25 March 1996 / Accepted: 13 September 1996     

干旱生境中接种丛枝菌根真菌对三叶鬼针草

为阐明丛枝菌根真菌对石灰岩地区适生植物三叶鬼针草(Bidens pilosa L.)光合作用的影响,设置正常浇水(A)、中度干旱胁迫(B)和重度干旱胁迫(C)3个水分处理梯度,比较了不同水分处理条件下接种丛枝菌根真菌Glomus mosseae和未接种三叶鬼针草之间净光合速率、气孔导度、蒸腾速率、胞间CO2浓度、羧化效率、水分利用效率等特征的差异.结果表明,水分胁迫显著降低三叶鬼针草的净光合速率、气孔导度、蒸腾速率和羧化效率.胞间CO2浓度在处理的前期(7d)因干旱胁迫而降低,在后期随土壤含水量的降低而升高;水分利用效率则是中度胁迫的植株、正常浇水处理植株、重度胁迫植株依次降低.在正常浇水条件下接种G. mosseae 对三叶鬼针草光合参数没有产生显著性影响;在中度胁迫条件下,接种植株较未接种植株在水分处理的前28d有更高的净光合速率、气孔导度、蒸腾速率和羧化效率;在重度胁迫条件下,虽然净光合速率、气孔导度、蒸腾速率和羧化效率接种植株高于未接种植株,但是二者并不显著.研究认为,干旱胁迫对三叶鬼针草光合作用的影响在水分处理的前期表现为气孔因素制约,在后期则主要是非气孔因素的影响;在正常浇水条件下接种G. mosseae 对三叶鬼针草的光合作用没有显著性影响,在干旱胁迫条件下,丛枝菌根真菌通过改善三叶鬼针草气孔导度和羧化效率等减弱干旱胁迫对植株的伤害,但是这种保护作用因为土壤水分的严重匮乏以及土壤干旱的时间延长而受到限制.