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.

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。

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.

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.

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。

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．

Assimilating efficiency of soil nutrition of Phalaris arundinacea L.

Phalaris arundinacea L. is fine perennial herbage of Gramineae. Eight treatments including N, P, K, NP, NK, NPK, PK and controls were done for studying its absorbing efficiency of soil nutrition. At tillering stage, the coefficient of absorbing and utilizing of N, P, and K from soil and fertilizer were analyzed according to M.K. Kaeomobk抯 formula and the contents of crude protein and fat were also measured. The yields of fresh crop, hay and seed were measured at ripening stage. For N absorbing coefficient of Ph. arundinacea from soil (Kn%), the composition PK treatment is best (12.36%) and K treatment is worse, while for N absorbing coefficient from fertilizer (Ky%), all the treatments are higher than control but the composition NPK treatment presented the highest (14.78%). For the absorbing coefficient of P from soil (Kn%), composition NK treatment is highest (19.30%), but K treatment was lower than control, while for that of P from fertilizer (Ky%), all the treatment were higher than control but the composition NP treatment showed the highest (19.52%). As to K absorbing coefficient from soil (Kn%), NP treatment is the best (19.2%) and single P treatment is worst, while for that (Ky%) of K from fertilizer the composition NPK treatment was best (28.44%). As concerns treatments for the outputs of fresh and dry crop, the composition NP treatment was best and all the treatment were higher than control. The composition NK treatment produced highest yield of seed and highest content of crude protein as compared to other treatments. For content of crude fat, the compositions NK and N treatments are best. The experiment indicates that application of fertilizer at proper proportions could increase the yield of Ph. Arundinacea and raise the utilizing efficiency of nourishment element from soil and fertilizer.

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.

NaCl与Na_2CO_3对高羊茅种子萌发及幼苗生长胁迫效应比较

分别用NaCl(50、100、200、300 mmol/L)和Na2CO3(5、10、15、25 mmol/L)对高羊茅(Festuca arundinacea Schreb)种子和幼苗进行胁迫处理,测定高羊茅种子发芽势、发芽率、幼苗根长、苗高、鲜重、叶绿素含量、根系脱氢酶活性、质膜透性和丙二醛含量等指标。结果表明:与对照相比,随着NaCl浓度的增加,高羊茅的发芽势、发芽率、幼苗根长呈下降趋势,叶质膜透性和根丙二醛含量呈上升趋势,苗高、鲜重、叶绿素含量和根系脱氢酶活性呈先上升后下降趋势,根质膜透性和叶丙二醛含量则呈先略下降后上升趋势;随着Na2CO3浓度的增加,其种子发芽势、发芽率、幼苗根长、苗高、鲜重、叶绿素含量均呈下降趋势,根和叶质膜透性和丙二醛含量均呈上升趋势,根系脱氢酶活性则呈先上升后下降趋势。综合分析表明,高羊茅对NaCl和Na2CO3的耐受浓度阈值分别为246.9 mmol/L和20.4 mmol/L,因此,高羊茅的耐盐性强于其耐碱性。

Trinexapac-ethyl、Prohexad ione-Ca在草坪化学控制中的应用研究

为探明Trinexapac-ethyl和Prohexadione-Ca用于草坪化学控制的可行性,研究了其对高羊茅(Festuca arundinacea)生长发育的影响.结果显示:使用剂量高于0.095 kg/hm2时, Trinexapac-ethyl对高羊茅具有明显的矮化作用,在0.36 kg/hm2剂量下Prohexadione-Ca的抑制率可达95.2%;Trinexapac-ethyl和Prohexadione-Ca都能使植株新增鲜重显著降低,但对新增干重影响很小,Trinexapac-ethyl对植株新增干重无影响;在施用Trinexapac-ethyl的情况下,随着留茬高度的增加新增株高减少,合理留茬高度为4～6 cm;Trinexapac-ethyl和Prohexadione-Ca可以有效降低株高,从而减少草坪修剪次数.

NaCl胁迫下高羊茅生长及K^+、Na^+吸收与运输的动态变化

通过室内水培试验研究探讨了NaCl胁迫对高羊茅苗期生长及K+、Na+吸收与运输的影响及其随时间的变化。结果表明,高羊茅苗期的干物质重和分蘖数随盐分水平的升高而降低,且与NaCl浓度存在极显著的负相关关系(r=-0.9902**);NaCl胁迫对高羊茅苗期茎叶生长的抑制作用大于对根系的影响;NaCl盐浓度的增加使高羊茅地上部干物质重、K+的相对含量、K+/Na+、SK+/Na+吸收降低,Na+的相对含量增高;胁迫时间对K+、Na+含量的影响显著。高羊茅根系对K+的吸收具有较高的选择性,吸收选择性系数SK+/Na+吸收主要受环境盐分浓度的影响,但运输选择性系数S++却主要受胁迫时间的影响,且盐分阈值随胁迫和生长时间的增加而呈下降趋势。

Ca(ClO)_2消毒剂对高羊茅种子的消毒效果

为研究Ca(Cl O)2对高羊茅种子作为外植体诱导培养愈伤组织时的消毒效果,以3种不同的前处理方式配合Ca(Cl O)2溶液不同浓度(3%、20%)和不同消毒时间(15、30、45、60 min)进行高羊茅种子消毒处理,以升汞消毒剂处理的高羊茅种子为对照。愈伤组织诱导培养结果表明:仅用Ca(Cl O)2搅拌的前处理的感菌率显著高于其余两种前处理;Ca(Cl O)2高浓度处理比低浓度处理的出愈率低;Ca(Cl O)2处理的出愈率和愈伤总数均优于升汞处理;最佳的消毒组合是无菌环境下用Ca(Cl O)2搅拌+3%Ca(Cl O)2浸泡60 min,该消毒处理方式可降低种子感菌率、缩短出愈时间、提高出愈系数,为建立高羊茅高效再生体系奠定实验基础。

Effects of Low Temperature-tolerant Microbial Inoculants from Municipal Solid Waste Compost on Turf Storage in Winter

Beneficial microorganisms were extracted from municipal solid waste compost and treated under low temperature stress to prepare complex microbial inoc- ulants. Turfgrass was inoculated with the prepared microbial inoculants to investigate the ecological effect of ~crobial inoculants on turf storage in winter. The re- suits showed that complex low temperature tolerant microbial inoculants significandy improved plant height and aboveground biomass of turfgrass. Compared with the non-inoculated control, plant height and biomass of the first batch of inoculated turfgrass were enhanced by 6.47% and 14.32%, respectively; and they were en- hanced by 6.94% and 17.41%, respectively, for the second batch of inoculated turfgrass. Under low temperature stress in winter, the reviving rate of inoculated tarfgrass was significandy higher than that of control, which was improved by 3.34% and 43.33% for the first and second batches of inoculated turfgrass respective- ly. After low temperature storage in winter, inoculated turfgrass revived in advance; specifically, the reviving date of the second batch of inoculated turfgrass was 3 d earlier than that of control. This low temperature tolerant microbial consortium was effective for turfgrass as an ecofriendly and acceptable technology to improve plant performance and development in winter and accelerate time to market of turf.

土壤基质填充废胶粒建植高羊茅对NaCl胁迫的生理响应

通过盆栽土壤基质填充废胶粒,研究草坪植物高羊茅对NaCl胁迫的生理响应。结果表明,填充胶粒能在一定程度上提高植物的地下生物量和叶绿素含量;当NaCl浓度超过0.4%时,与对照相比胶粒处理地上生物量、地下生物量有所增加。草坪基质中填充胶粒对过氧化物酶(POD)活性没有显著影响,但NaCl浓度高于0.4%时,胶粒处理POD均高于对照;在所设的NaCl浓度范围内,胶粒的加入明显增加了超氧化物歧化酶(SOD)活性,尤其在0.2%NaCl浓度下SOD活性比对照提高了161.66%。另外,胶粒能显著缓解NaCl胁迫导致的高羊茅叶片丙二醛(MDA)含量和相对电导率的增加,对脯氨酸含量的影响不大。在草坪基质中填充废胶粒,可以减轻盐胁迫对高羊茅的伤害,提高高羊茅的抗盐性。

高羊茅黔草1号和黔草2号LEA3基因克隆与苗期干旱胁迫表达

【目的】克隆贵州本地高羊茅品种黔草1号和黔草2号的第三组晚期胚胎发生丰富蛋白(Late Embryogenesis Abundant Protein,Group 3,LEA3)基因,分析其生物信息学及其苗期干旱胁迫的表达量变化,为进一步探讨LEA3基因的功能和高羊茅抗旱分子机制提供理论基础。【方法】参照NCBI已登录大麦LEA3基因序列,利用RT-PCR获得黔草1号和黔草2号LEA3基因cDNA序列,应用相关软件对氨基酸序列进行分析,采用实时荧光定量PCR(qRT-PCR)分析苗期干旱胁迫的表达量。【结果】黔草1号和黔草2号LEA3基因完整的ORF框长度分别为609和642 bp,分别编码203个氨基酸和213个氨基酸;黔草1号LEA3蛋白比黔草2号缺失1个基元序列(包含11个氨基酸),且两者LEA3基因编码的氨基酸序列存在12个位点的氨基酸差异;黔草1号LEA3基因编码蛋白分子量为20.72 kDa,等电点8.55,总平均亲水性为-0.971;黔草2号LEA3基因编码蛋白分子量为22.01 kDa,等电点8.99,总平均亲水性为-1.069;两者的LEA3蛋白均没有跨膜结构,且均以α-螺旋结构占主导,与大麦、山羊草、冰草和小麦的LEA3具有高度的同源性,与小麦、旱麦草、大麦、山羊草、无芒雀麦、冰草的LEA3蛋白聚为一类;苗期干旱胁迫下黔草1号LEA3基因的表达量在整个胁迫过程中总体高于黔草2号,2个品种高羊茅在-2.5 MPa PEG 6000胁迫下的LEA3基因表达量高于-1.5 MPa PEG 6000胁迫。【结论】黔草1号和黔草2号的抗旱性与LEA3基因的表达量呈正相关,相同干旱胁迫条件下,黔草1号LEA3的表达量高于黔草2号,推测在受到干旱胁迫时,黔草1号更能适应干旱环境。

NaCl与Na_2CO_3对高羊茅生理胁迫的差异分析

分别采用浓度为50～250mmol/L的中性盐(NaCl)和浓度为25～125mmol/L的碱性盐(Na2CO3)对生长7d的高羊茅幼苗进行胁迫处理,测定并分析了NaCl与Na2CO3对高羊茅含水量、可溶性蛋白、可溶性糖、叶绿素含量及SOD、POD活性等生理生化指标的影响。结果表明:随盐浓度升高,高羊茅幼苗的含水量和SOD活性总体呈下降趋势,少数浓度时指标也有升高;可溶性蛋白、可溶性糖、叶绿素含量及POD活性随盐浓度增加而逐渐升高。50mmol/L Na+浓度时,两种盐对含水量、可溶性蛋白、可溶性糖及POD活性的影响相似;随处理浓度的升高,碱性盐对高羊茅各项生理生化指标的影响大多比中性盐的影响要大,表明高羊茅对中性盐的耐受力比碱性盐要强。

Study on Composition of Culture Medium for Indoor Ecological Lawn

The compositions of soilless culture media for indoor lawns were studied through the determination of quality indexes （grass height, clipping weight, leaf width, density, lawn color and uniformity） of Festuca rubra L. and Perennial Lolzum perenne L lawns. The results showed that composition 4 （black turf 1.2 L＋vermiculite 0.3 L＋ceramsite 0 L） is not suitable for the cultivation of indoor lawns. Composition 1 （black turf 0.3 L＋vermiculite 0.75 L＋ceramsite 0.45 L） and composition 2 （black turf 0.6 L＋vermiculite 0.6 L＋ceramsite 0.3 L） are suitable for indoor cultivation of F. rubra; and Perennial L. perenne turned yellow in all the tested media, and therefore, the media tested in this study are not suitable for indoor cultivation of Perennial L. perenne.