禾本科植物内生真菌研究14:Neotyphodium sinofestucae在小颖羊茅体内的分布及其种传特性

2010年在南京紫金山地区采集小颖羊茅(Festuca parvigluma Steud.)的完整植株和成熟种子,通过染色、分离、特异性基因片段检测等方法探究内生真菌Neotyphodium sinofestucae Chen,Ji et Wang在其体内的分布和种传特性。从植株茎秆、叶鞘、叶片和种子稃片及颖果等部位分别检测和分离到具有禾本科植物内生真菌特征的真菌。同时,从含有内生真菌的种子培育实生苗,并从实生苗中检测和分离到类似的内生真菌。通过对tubB基因片段的PCR扩增和检测,证明分离自小颖羊茅的样品与其实生苗的分离菌株是相同的N.sinofestucae。上述结果表明:N.sinofestucae遍布小颖羊茅植株地上部分,却不进入根部;内生真菌N.sinofestucae能进入种子,通过种子进行垂直传播,与宿主植物长期互利共生。

盐胁迫对高羊茅(Festuca arundinacea)幼苗生长和离子分布的影响

盐胁迫环境抑制植物的生长,影响植物组织的离子分布,不同的盐分组成对植物的抑制伤害存在差异,为了研究上海市临港新城滨海盐渍土的生态恢复和重建,模拟该地区的盐分组成,进行了高羊茅(Festuca arundinacea)幼苗的盐胁迫试验。高羊茅种子在非盐胁迫条件下萌发,出苗5d后,进行了不同浓度NaCl:0、50、100、150、200、300、400mmol/L处理,15d后测定生长情况、组织含水量和Na+、K+、Ca2+、Mg2+等离子含量。研究结果表明:盐分对高羊茅幼苗的抑制作用随NaCl浓度增加而加剧,低盐胁迫环境下,幼苗地上部分和根系的鲜重、干重和含水量都与对照没有显著性差异,但是高盐环境严重影响了高羊茅幼苗的生长,而且对地上部分的抑制作用大于根部;盐胁迫影响植物组织的离子分布,Na+浓度持续增加,Ca2+和K+浓度下降,Mg2+含量的影响不大;各组织中K/Na、Ca/Na和Mg/Na随盐胁迫增加而下降。

分离自苇状羊茅(Festuca arundinacea Schreb.)的内生真菌Neotyphodium uncinatum

从苇状羊茅植株中发现了与Neotyphodiumcoenophialum不同的内生真菌 ,经过分离、培养 ,分离菌株被鉴定为Neotyphodiumuncinatum (W .Gams ,Petrini&D .Schmidt)Glenn ,Bacon&Hanlin。通过对苇状羊茅植株各部位的调查 ,确认该菌在植株的地上部分有较为系统的分布。所发现的Neotyphodiumuncinatum

高羊茅(Festuca arundinacea)对土壤中菲和芘的修复作用研究

采用盆栽试验法，研究了高羊茅对污染土壤中菲、芘的去除效果与修复机制．结果显示：在试验浓度范围（0～322mg／kg）内，植物-微生物系统对土壤中芘、菲的去除效果明显．种植高羊茅60d后，土壤-高羊茅系统（TD3）对土壤中菲、芘去除率分别为52．82％～82．27％（m=67．96％）、47．27％～75．4％（m=60．06％）；平均去除率分别比对照组TD1（无植物，不加NaN3）高45．41％，41．63％，比对照组TD2（无植物，加0．1％NaN3）高62．89％，57．5％．高羊茅能吸收积累土壤中的菲和芘，根部和茎叶部菲、芘含量随土壤中菲、芘浓度的提高而增大；生物浓缩系数随土壤中菲、芘含量的增加而减小，且根部大于茎叶部、芘大于菲．修复过程中，非生物因子、植物积累对菲、芘的去除率分别只有5．07％，0．017％和2．56％，0．11％；微生物降解、高羊茅-微生物间的交互作用对菲、芘的平均去除率分别为19．21％，62．87％和15．87％，57．5％．说明微生物降解作用、植物-微生物交互作用是土壤中菲、芘去除的主要途径．

Cloning and Expression Analysis of FaSRP Gene in Festuca arundinacea under Abiotic Stresses

[Objective] This study aimed to study the structure and functions of SRP gene and variation in its expression under abiotic stresses. [Method] Using the SRP sequence obtained from transcriptome sequencing as the template, the full-length cDNA sequence of SRP gene in Festuca arundinacea was amplified using the 3＇RACE and 5＇RACE methods. [Result] The cDNA sequence of SRP gene has a full length of 1 165 bp, and it contains an open reading frame in full length of 855 bp. The encoded protein by SRP gene is composed of 284 amino acids, and contains a REF domain. The bioinformatic researches on structures and functions of SRPs show that the SRP gene in Festuca arundinacea （FaSRP） has relatively high ho- mologies with SRPs in monocots. Under low nitrogen, drought, high temperature and high salt stresses, the variations in expression of FaSRP gene were studied using fluorescence quantitative PCR. The results showed that FaSRP gene makes responses to low nitrogen, drought and high temperature stresses, but the relevant response mechanisms are not the same, indicating different pathways regulating re- sistance of plants. The expression of FaSRP gene is insensitive to high salt stress. [Conclusion] This study will provide certain candidate gene and technical reserve for breeding of drought- and high temperature-tolerant, nutritious and highly efficient Festuca arundinacea cultivars.

Physiological Response of Space Flight Mutation New Strains of Festuca arundinacea to High Temperature Stress and Comprehensive Evaluation

[Objective] The paper was to understand the physiological response of space flight mutation new strains of Festuca arundinacea to high temperature stress. [Method] The influence of high temperature stress on eco-physiological characteristics of 11 F. arundinacea materials was studied in a pot experiment.Physiological and biochemical indexes,including soluble protein content,superoxide dismutase( SOD),peroxidase( POD) and catalase( CAT) activities,were measured and analyzed once every 4 d for a total of three times. Subordinate function and grey rational analysis were used to comprehensively evaluate the heat resistance of 11 materials. [Result] The soluble protein content decreased with the extension of heat resistance stress,while SOD,POD and CAT activities showed an upward trend. The high temperature resistance of 11 F. arundinacea materials was evaluated by subordinate function method. The resistance order was SP5-85> SP5-60 > SP5-71 > SP5-42 > SP5-94 > SP5-7 > SP5-88 > F. arundinacea cv Shuicheng > SP5-5 > SP5-89 = F. arundinacea cv Qiancao No. 1. The relational order of various heat resistance indexes and heat resistance obtained by grey rational analysis was SOD > CAT > POD > soluble protein content. [Conclusion]The result provides a theoretical basis for stress physiology and stress breeding of cold season grass in southwest region.

Cloning and Bioinformatics Analysis of FaPHYC Gene in Tall Fescue (Festuca arundinacea)

The cDNA of PHYC gene of tall fescue ( Festuca arundinacea ) was cloned by RT-PCR,named FaPHYC.The sequence was analyzed by bioinformatic software,and the results showed that the full-length cDNA(3816 bp) was obtained with an open reading frame predicated to be 3 411 bp,which encoded 1 136 amino acids.By BLASTP analysis,the N-terminal of FaPHYC was composed of GAF and pPhytochrome domains,and its C-terminal included two repeated PAS domains,one histidine kinase A domain and one histidine kinase-like ATPase domain.The phylogenetic analysis showed that FaPHYC was closely related to PHYCs from other Gramineae plants.The qRT-PCR results showed that FaPHYC expression was regulated by drought stress,heat stress,salt stress and nitrogen stress.The real-time quantitative PCR assay showed the expression of FaPHYC in tall fescue seedlings was continuously up-regulated during the light period and down-regulated in the dark.The results suggest that FaPHYC is photoperiod sensitive and plays a functional role in flowering regulation by photoperiod in tall fescue.The FaPHYC gene was cloned and reported from tall fescue for the first time,which lays a foundation for the investigation of the stress mechanism of FaPHYC in tall fescue plant.

Seedling Blight of Festuca arizonica Caused by Rhizoctonia solani

We report here a damping-off disease occurring at the seedling stage of the grass Festuca arizonica (Arizona fescue) that is caused by the soil-borne fungal pathogen Rhizoctonia solani.

Effects of Microbial Inoculants from Sewage Sludge on Initial Growth of Festuca arundinacea L.

Microbial inoculants were isolated from sewage sludge and applied to turfgrass medium. Effects of microbial inoculants on seed germination and initial growth of Festuca arundinacea L. were analyzed to investigate the optimal proportion of compound microbial inoculants. Significant differences were found in various growth indices and chlorophyll content of F. arundinacea between microbial treatment and control The combination of Pseudomonas stutzeri and Trichoderma reesei exhibited the best results. Compared with the control, seed germination rate, seedling height, shoot dry weight, root weight, chlorophyll a content and chlorophyll b content of F. arundinacea treated with the combination of Pseudomonas stutzeri and Trichoderma reesei were improved by 11.60%, 43.47%, 148%, 159%, 144% and 136%, respectively. Based on comprehensive analysis of various indicators, the optimal proportion （V/V） of microbial combination was Pseudomonas stutzeri： Tfichoderma reesei = 1： 1, which was the most conducive to plant growth and development.

Damping-Off of Festuca arizonica Caused by Fusarium

We report here a damping-off disease occurring at the seedling stage, resulting in severe loss of the seedling stands caused by an important soil-borne fungal pathogen, Fusarium oxysporum. Endophyte-infected plants appear to be more resistant to the damping-off disease than uninfected plants.

Advances in Breeding of Festuca arundinacea Schreb.

In this paper, domestic and foreign advances in breeding situation and conventional breeding methods of Festuca arundinacea Schreb. and the applica- tion of modem biological techniques （ somaclonal variation, somatic cell hybridization, genetic map construction, genetic diversity, gene cloning and expression, ge- netic engineering） in breeding of new varieties of F. arundinacea were summarized, which proposed new ideas for the development of F. arundinacea breeding and provided reference for breeding of new varieties of F. arundinacea.

Seed Protein Electrophoresis for Assessment of Genetic Diversity among Festuca pratensis Huds, Local Ecotypes

Fifty two accessions of Festuca pratensis Huds. were analysed of which 18 had been collected in Switzerland and 32 in Bulgaria. SDS-PAGE of seed proteins was implemented to detect genetic variation among ecotype populations using a modification of UPOV method for barley. The modification concerns mainly protein extracting procedure as detailed described in the protocol. The two cultivars Preval and Cosmolit were used as standards. Cluster analyses and correspondence analyses/scatter plot were used as statistic approaches for determining genetic diversity among individual ecotypes and groups of ecotypes. Electrophoretic spectra of proteins show clear differences among local accessions in relation to their origin. In Swiss ecotypes 32 protein fragments were determined whereas in Bulgarian local populations their number was 68. Each of the two eco-groups possesses fragments that appear in all accessions of the group. The number of monomorphic bands within Bulgarian local ecotypes is four whereas their number in Swiss ecotypes is 12. Four monomorphic bands appearing in all proteinograms no difference of eco-groups was identified with Rm values of： 0.43, 0.55, 0.58, 0.82. A higher level of protein band polymorphism was proven in Bulgarian ecotypes in comparison with Swiss ecotypes. Thirty seven polymorphic bands occurred exclusively in the Bulgarian local ecotypes and had a frequency of 0.03 or higher whereas within Swiss ecotypes was detected one unique protein fragment. SDS-PAGE ＂fingerprinting＂ is suggested as a fast and easy approach to differentiate F. pratensis ecotypes by their origin as well for detection of foreign germplasm for inclusion in breeding programs.

热锻炼对高羊茅(Festuca arundinacea)和多年生黑麦草(Lolium perenne)抗高温能力的影响

夏季高温胁迫已成为限制冷季型草坪草生长和发育的一个主要问题。以两种耐热性不同的冷季型草坪草高羊茅和多年生黑麦草(前者较耐热)为材料,经过3d 30℃的热锻炼预处理后,分别在38、42、46℃的高温下处理14h。在这些高温条件下,研究了经过热锻炼预处理的高羊茅和多年生黑麦草叶片膜脂过氧化、抗氧化剂含量以及叶绿体超微结构的变化。结果表明:(1)热锻炼提高了高羊茅和多年生黑麦草的耐热性,显著缓减了高温条件下两种草坪草叶片膜脂过氧化程度的加剧,降低了叶片过氧化氢(H2O2)和超氧阴离子(O2.-)的产生速率。(2)高温条件下,热锻炼使高羊茅和多年生黑麦草叶片中抗氧化剂抗坏血酸(AsA)和谷胱甘肽(GSH)的含量下降程度有所缓减。(3)热锻炼减轻了高温胁迫对高羊茅和多年生黑麦草叶片叶绿体超微结构的损伤。这些结果说明热锻炼能够减轻高温对草坪草叶绿体的伤害可能与其在高温胁迫下和对照相比具有较高的抗氧化剂含量有关,这也可能是冷季型草坪草对高温的适应机制之一。

Comparative Study on the Effects of Three Carbon Nanomaterials on the Growth of Festuca arundinacea in Compost Substrate

[Objectives] This study was conducted to provide a scientific basis for the utilization of municipal solid waste resources, the remediation of contaminated substrates and the ecological safety of urban lawn planting. [Methods] Graphene, multi-layer graphene oxide and multi-walled carbon nanotubes were added to municipal solid waste(MSW) compost as turf substrate to determine the morphological indicators such as turfgrass biomass, chlorophyll contents and plant height. [Results] There were no significant differences in the plant height of Festuca arundinacea between different treatment groups in the first 30 d. The effects of adding different carbon nanomaterials on the fresh and dry weights of F. arundinacea were not significantly different. The aboveground biomass of F. arundinacea was the largest after adding graphene oxide, and the underground fresh weight decreased significantly in the hydroxyl multi-walled carbon nanotube treatment compared with the control. As to the chlorophyll content, the graphene oxide treatment was the highest, but there were no significant differences between all the treatment groups and the control group. [Conclusions] This study can provide data support for MSW compost substrate, lawn planting system and heavy metal pollutant passivating agents.

Study on Sterilization Method of Endophytic Fungi in Tall Fescue(Festuca arundinacea)and Ryegrass(Lolium perenne)Seeds

[Objectives]This study was conducted to obtain the best solution for sterilizing tall fescue(Festuca arundinacea)and ryegrass(Lolium perenne)seeds by heating in a water bath for a short time.[Methods]The tall fescue and ryegrass seeds infected with endophytic fungi were sterilized by heating in a water bath to compare seed germination and initial growth of turfgrass seedlings under different treatment time.[Results]Sterilization in a 60℃water bath for 20 and 30 min both inhibited the germination of tall fescue and ryegrass seeds,while the 20 min treatment did not significantly affect the lengths of stems,leaves and roots of seedlings.The 20 min water bath sterilization treatment had no effects on the plant heights and biomass of the two turfgrass seedlings and the tiller number of ryegrass,but sterilization for 30 min significantly reduced the aboveground and total biomass of seedlings and the tiller number of ryegrass.[Conclusions]Sterilization in a water bath at 60℃ for 20 min achieved rapid sterilization in a short time,without significantly negatively affecting the growth of seedlings.

Morphological Feature and Physicochemical Characteristics of Soils under Festuca spp. Dominant Steppe at High Mountain and Mountain of Khuvsgul, Mongolia

The morphology, physicochemical, humic substances and micromorphological characteristics of four soil profiles of the steppe dominant by Festuca lenensis (F. lenensis) at the high mountain and mountain of Khuvsgul, Mongolia were studied. Soils were classified as Regosols and Leptosols at high mountain steppe, Leptosols and Cambisols at mountain steppe. On a high mountain, the plant root distribution, OC, N and moisture contents were high due to its high precipitation and low temperature. The soils show immature characteristics with low available nutrients, weakly developed crumb structure, many semi- and undecomposed plant residues, and few little organic pigments with few excrements. The humic acids with immature to degraded characteristics indicate that the climatic condition of high mountains inhibits the soil decomposition process. Due to extremely different landform positions, there a sharp difference was observed between studied soils on high mountain steppe. On the summit with a flat position, the soil of TSO1 showed finer soil texture with higher CEC, exchangeable Mg2+ and humification degree of SOM compared with the soil of TSO2, which located on the steep slope. This confirms that the abrupt changes in landform on high mountain strongly affect the properties of topsoil. On the mountain steppe, the soil contains higher exchangeable Na+, exchangeable K+ and water soluble at topsoil;however, the plant root distribution, OC, N and moisture contents were lower than that of high mountain soil. Because of warmer air and soil temperature in comparison with that of high elevation, active turnover in humic horizon and chemical weathering process lead to higher available nutrients in mountain steppe. The degraded to well humified characteristics of humic acid, moderately developed crumb structure, a higher component of little organic pigment and many intact excrements indicate that the soil decomposition process and biological activity were higher than that of the high mountain steppe. Our finding suggests that the climatic condition dependent on altitude and landform position at the high mountain and mountain of Khuvsgul had a large impact and played a key role in the soil properties and characteristics of steppe dominant by F. lenensis.

6个苇状羊茅(型)品种在成都平原区的生产性能评价

为筛选适宜成都平原推广利用的优异牧草品种,对6个苇状羊茅(型)品种进行了生育期、主要农艺性状、鲜(干)草产量及牧草营养成分等指标的测定,并运用灰色关联度分析进行综合评价。结果表明,‘特沃’和‘科瑞’的生育期最长,均达275 d。各参试品种平均再生速度依次为‘马胡丽娜’>‘科瑞’>‘都脉’>‘特沃’>‘K31’>‘福泰’。品种‘马胡丽娜’年平均干草产量最高,可达12790.99 kg·hm-2,分别较国审品种‘特沃’和‘都脉’显著增加14.62%和4.28%(P<0.05),表现出明显的产量优势。‘科瑞’干草产量较‘特沃’显著增加6.59%(P<0.05),且植株自然高度可达100.15 cm,高于‘特沃’和‘都脉’(P<0.05)。‘都脉’的分蘖数最大,为47.33个·株^(-1),显著高于其他品种(P<0.05),其次是‘科瑞’和‘马胡丽娜’,分别达37.50和37.00个·株^(-1),较品种‘特沃’多6.25和5.75个·株^(-1)。‘K31’茎叶比为0.96,高于其他参试品种。对6个参试品种的营养成分测定表明,‘特沃’的粗蛋白含量最高,可达18.60%,‘科瑞’的粗脂肪含量最高,可达5.41%。通过灰色关联度法综合比较分析,‘科瑞’表现最优,‘特沃’和‘马胡丽娜’综合性状表现较优,适宜在成都平原地区推广种植利用。

三叶草根浸提液对3种牧草种子萌发及幼苗生长的影响

为探索三叶草的防控及利用提供理论依据,采用培养皿生物测定法分析,研究不同浓度(0、2.5、5.0、10.0 g·L-1)三叶草根浸提液对黑麦草(Lolium perenne)、高羊茅(Festuca arundinac)和赖草(Leymus secalinus)种子萌发及幼苗生长的化感作用,以及三叶草(Trifolium repens)的生长竞争力和成功入侵原因。结果表明,三叶草根浸提液对3种牧草种子萌发(发芽率、发芽指数)和幼苗生长(根长、苗长、根干重、苗干重)均有化感作用,且其化感作用效应存在一定的差异。三叶草根浸提液对黑麦草种子萌发具有抑制作用,对高羊茅和赖草的种子萌发有“低促高抑”作用,对黑麦草根长和干重、苗高和干重均具有抑制作用,对高羊茅和赖草则表现为低浓度的促进、高浓度的抑制效应。从综合化感效应指数来看,随着三叶草根浸提液浓度的增加,对黑麦草具有抑制作用,而对高羊茅和赖草有“低促高抑”的双重作用。3种供试牧草对三叶草根浸提液的敏感程度也有差异,表现为:黑麦草>高羊茅>赖草。入侵植物三叶草根浸提液对3种牧草的化感效应较为明显,在入侵严重的地方,可选择补播耐受力强的牧草。

7份羊茅属种质芽期抗旱性评价

为了探究羊茅属种质芽期对逆境的胁迫能力,以沟羊茅新品系(FV)与紫羊茅常用品种传奇(CQ)、草原(CY)、梦神(MS)、北方(BF)、玉山(YS)和多花(DH)为材料,采用不同浓度PEG-6000(0、5%、10%、15%、20%、25%)模拟干旱胁迫,在芽期测定了发芽率、发芽势、活力指数、胚根长、胚芽长、根芽比等萌发指标,分析干旱对其种子萌发的影响,并结合主成分分析、隶属函数分析、聚类分析等对其抗旱性进行评价。结果表明,高浓度PEG(20%~25%)显著降低了羊茅属种质的发芽势、发芽率、发芽指数、活力指数、胚根长、胚根长及根芽比,而5%~15%PEG则因种质的差异而变化趋势有所不同,其中0~10%PEG对FV、BF、YS的发芽率、发芽势、活力指数及发芽指数具有显著的引发作用;综合评价表明,主成分分析将7个相对单项性状指标降维成2个新的综合指标,累积贡献率达85.06%;以新合成的综合指标为依据,采用隶属函数法结合聚类分析,将7份羊茅属种质分为三类,其中,FV、CQ和BF的抗旱能力较强,CY及YS的抗旱能力一般,而MS、DH的抗旱能力较弱。

施钾对苇状羊茅生长、生理特性及钾吸收的影响

【目的】明确钾肥施用对苇状羊茅(Festuca arundinacea)生长、生理特性的影响,确定苇状羊茅适宜的钾肥用量和钾营养诊断指标。【方法】通过盆栽试验,研究苇状羊茅地上部生长、生理和钾素养分吸收对不同钾肥用量(K_(2)O分别为0、18、36、54 g/m^(2))的响应。【结果】随钾肥用量的增加,苇状羊茅各生育阶段的株高和地上部干物质量均呈先升后降的变化趋势,钾肥用量36 g/m^(2)时达最大值;出苗后50~137 d和0~50 d植株株高增量和干物质累积速率较大。施用钾肥36 g/m^(2)有利于促进植株地上部钾的吸收,提高植株叶片的光合色素含量、抗氧化酶(POD)活性和可溶性蛋白含量,降低丙二醛(MDA)含量。增加钾肥用量提高了苇状羊茅栽培体系的钾素损失风险。适宜苇状羊茅生长的钾肥用量范围为K_(2)O 7.0~49.9 g/m^(2),获得80%最大地上部干物质量的地上部钾临界浓度为10.8 g/kg。【结论】钾肥施用对苇状羊茅生长和生理特性影响显著,钾肥用量7.0 g/m^(2)和地上部钾浓度10.8 g/kg可分别作为苇状养护管理的钾肥用量推荐和植株钾营养诊断参考。