Stable Transformation of Three Cultivars of Kentucky Bluegrass (Poa pratensis L.) by Particle Bombardment of Mature Seed-Derived Highly Regenerative Callus

High embryogenic calli of three cutivars of Kentucky bluegrass, Md, Bd, and Gm, were induced from mature embryos, and were proliferated on medium K3 and K5. Embryogenic calli were transformed with plamids pDM803 and pBY520 by microprojectile bombardment. Fourty-two transgenic lines had been obtained. The highest efficiency of transformation reached to 3.7% for cv. Md, 2.8% for cv. Gm, and 5 % for cv. Bd. The micro nutriment of Cupric had significant effect on transformation. The embryogenic callus cultured in dim-light condition had higher transformation efficiency than the green callus cultured in light condition for one month before transformation. The selective regime and selective pressure on the putative transgenic plants were important for obtaining the desire number of transgenic plants. It also affected the copy number of integrated genes in the genomic DNA of transgenic plants.

Growth of Kentucky Bluegrass as Influenced by Nitrogen and Trinexapacethyl

The experiment aimed to evaluate the effects of N and trinexapac-ethyl （TE） to the growth of Kentucky bluegrass. By using the method of randomized blocks design, 4 N levels, 0 （N1）, 20 （N2）, 25 （N3）, and 30 （N4） kg N ha t mon% with TE at 0 （T1）, 312.5 （T2） and 625 （T3） mL ha-1 every 10 d effect on the total height, clippings and chlorophyll concentration of Kentucky bluegrass were evaluated. Results showed that total height and total clippings of Kentucky bluegrass were reduced by treating with TE and increased by treating with N. Kentucky bluegrass treated with 312.5 and 625 mL ha ＇ per 10 d had total clippings and total height reduced 30 and 45% under different N application level. The N × TE interaction was significant for the chlorophyll concentration of Kentucky bluegrass. The chlorophyll concentration of Kentucky bluegrass treated with TE at 625 mL ha-1 every 10 d was significantly higher than that of Kentucky bluegrass treated with 312.5 mL ha-1 every 10 d at the same N level. The chlorophyll concentration of turfgrass treated with TE at 625 mL ha-1 every 10 d receiving 30 kg N ha -1 month-1 was the highest. Based on the index of turf growth and the chlorophyll concentration, turfgrass receiving 20 or 30 kg N ha-1 month-1 with 625 mL TE ha i every 10 d could balance the problem of mowing frequency and growth quickly while keeping a good quality.

Study on Classification and Genetic Diversity of Kentucky Bluegrasses by Using RAPD Markers

A total of 69 random primers were screened by using random amplified polymorphic DNA （RAPD） markers to analyze the genetic bands of 32 Kentucky bluegrass cultivars. A total of 197 bands were amplified from 46 primers, among which 195 bands were polymorphic. Each primer could amplify one to nine polymorphic bands with an average of 4.3 per primer. Based on similarity coefficient analysis of RAPD results and by using NTSYS software to cluster analyze with the average UPGMA method, the result showed that 18 cultivars of the 32 were in group 1, three cultivars were in group 2, two cultivars were in group 3, eight cultivars were in group 4, and only one cultivar in group 5.

Mowing Height and Mowing Frequency Interactions on Turf Performance of Kentucky Bluegrass

The mowing height and mowing frequency are the key factors that affect the turf quality and turfgrass growing ability. The purpose of this study was to reveal the alternative effcient interaction between mowing height and mowing frequency in Kentucky bluegrass turf management. The Kentucky bluegrass cultivar ＇Kenblue＇ （Poa pratensis L.） was used to evaluate the interactions of mowing heights （H1, H4 and H7） and mowing frequencies （F2, F5 and F8） on the performance of turfgrass and turf physiological responses under the feld condition. The results showed that the treatment of H7×F8 had perfect infuence on the turf texture, root elongation, and also enhanced soluble sugar and free proline content in plants. While, the turf quality, color, uniformity and the increasing tiller numbers were performed more superior in the treatment of H4×F5 than other treatments. In all the interacting treatments, the relation between leaf green index and number of shoots （dm2） was positively correlated, thus, it could be speculated that the proper mowing height with frequency had a strong impact on the increasing of turfgrass tillers. On the whole, the interaction of H4×F5 on turf quality was the most excellent treatment in this experiment.

Classification and Genetic Diversity of Kentucky Bluegrasses by Using RAPD Markers

Total 69 random primers were screened out through RAPD （Random Amplified Polymorphic DNA） markers to analyze the genetic bands of 32 Kentucky bluegrass cultivars. A total of 197 bands were amplified from 46 primers of them, among which 195 bands were polymorphic. Each primer could amplify one to nine polymorphic bands with an average of 4.3 being amplified from an average of primer. Based on the similarity coefficient analysis of RAPD results, NTSYS software and cluster analysis by using average UPGMA method, the result showed that 18 of 32 cultivars were in group 1, three cultivars were in group 2, two cultivars were in group 3, eight cultivars were in group 4, and there was only one cultivar in group 5, which was C2.

Callus induction and plant regeneration from mature bluegrass (Poa pratensis L.) seeds

A protocol was discussed for high efficient plant regeneration from seven bluegrass （Poa pratensis L.） cultivars via an in- direct callus induction and somatic embryogenesis method. Mature seeds were used as explants for callus initiation. Callus induction and proliferation efficiencies were investigated on NB, modified MS （MMS） and MS media, supplemented with 2.0 mg·L^-1 2,4-dichlorophenoxyacetic acid （2,4-D）. The MMS medium performed best. Based on the MMS medium, direct and indirect callus induction effects of bluegrass from mature seeds were compared at the range of 1-5 mg·L^-1 2,4-D contained in the medium. Under the direct callus induction method, the most suitable 2,4-D concentrations varied among cultivars. Under the indirect callus induction method, a significantly high callus induction frequency （93.33%-98.33%） was obtained and there were barely any statistically sig- nificant differences among the tested genetically diverse cultivars. Somatic embryos were promoted on the MMS medium supple- mented with 3 mg·L^-1 2,4-D, 0.1 mg·L^-1 kinetin and 0.8 mg·L^-1 CuSO4. Embryogenetic calli developed into plantlets on the MMS medium containing different concentrations of thidiazuron （TDZ）, and the differentiation frequencies varied in the range from 20.15% to 77.65%. The 0.25 mg·L^-1 TDZ was generally the most suitable concentration for the tested cultivars.

草地早熟禾BBML基因的克隆及表达模式分析

为探究婴儿潮(BABY BOOM,BBM)基因诱导草地早熟禾(Poa pratensis)体胚发生的潜在功能,本研究采用同源克隆方法获得PpBBML(PpBBM-like)基因,并对其进行生信分析、亚细胞定位和表达模式分析。结果表明:PpBBML编码的蛋白具BBM特有的bbm-1基序且定位于细胞核,并与小麦(Triticum aestivum)BBM亲缘关系最近。PpBBML的表达水平存在组织特异性,表达量为花药>茎基>幼叶>根茎>老叶>不定根。6-BA,NAA(除0 h外),MeJA和EBR(除4 h外)处理后的PpBBML表达量均高于CK;GA3处理后的PpBBML表达量均低于CK。在仅添加生长素的愈伤组织诱导培养基上,其表达量在接种第2 d最高。综上所述,PpBBML是AP2亚家族的转录因子基因,其表达量在幼嫩组织中最高且受到不同激素的诱导,并在愈伤组织形成初期表达量急剧上升,因此,该基因可能具有诱导草地早熟禾胚性愈伤组织发生的潜力。

草地早熟禾胚胎发生标记基因BBM的鉴定及系统进化分析

【目的】利用生物信息学方法,鉴定草地早熟禾(Poa pratensis)胚胎发生标记基因PpBBM,并通过保守基序比对和进化树分析,推测可能具有孤雌生殖功能的PpBBM基因。【方法】基于草地早熟禾转录组数据库信息,检索PpBBM基因。通过生物信息学软件分析PpBBMs蛋白理化特性,比较不同PpBBMs蛋白的系统进化关系和保守基序差异。【结果】共获得3个具有完整开放阅读框的PpBBMs基因,其编码的蛋白均含有2个AP2结构域,其中,PpBBM1和PpBBM2具有相同的motif类型和数量。3个PpBBMs蛋白都是酸性蛋白,定位于细胞核,不存在跨膜结构和信号肽。系统进化分析表明,30个物种的BBM/BBML蛋白成员共聚为5类。其中,PpBBM1、PpBBM2聚在第2分支,分别与BdBBM和SbBBML亲缘关系最近;PpBBML位于第5分支,与ZmBBML亲缘关系最近。PpBBM1、PpBBM2和PpBBML均具有BBM基因所特有的bbm-1基序以及AP2亚家族特定的euANT2-4基序,但PpBBML蛋白的所有基序均有个别氨基酸残基存在变异。【结论】与PpBBML基因相比,PpBBM1和PpBBM2更有可能具有诱导孤雌生殖的功能。

补光光源和日累积光照量对草地早熟禾草坪质量的影响

为降低体育场遮荫环境对草坪草生长的制约,提高运动场草坪的质量与性能,本研究以不补光为对照(CK),选用发光二极管(LED)和高压钠灯(HPS)作为补光光源,分别在11.18,15.12和18.36 mol·(m^(2)·d)^(-1)的日累积光照量(Daily light integral,DLI)下培育草地早熟禾(Poa pratensis)草坪56天,探讨了补光对草坪温湿度、草坪质量、生物量、根系生长和叶绿素含量的影响。结果表明:HPS补光对草坪冠层增温效果高于LED,同时HPS补光降低了冠层湿度。LED和HPS补光灯均能维持草地早熟禾的草坪质量,增加DLI有利于草地早熟禾的生物量累积和根系生长,在DLI为18.36 mol·(m^(2)·d)^(-1)的处理下效果最佳。在LED补光18.36 mol·(m^(2)·d)^(-1)下生长的草地早熟禾的叶绿素a和叶绿素b含量显著高于HPS补光(P<0.05)。因此,LED和HPS均可应用于运动场草坪补光,建议设置DLI为18.36 mol·(m^(2)·d)^(-1)进行草地早熟禾草坪的补光养护,在夏季或气温较高的时候推荐使用LED补光。

传统蓝染的植物和染法及其工艺简史

蓝染是一种传统工艺,适用于不同社会阶层与各种纤维染色,也是世界许多民族的重要文化表现形式。蓼蓝、木蓝、菘蓝、马蓝(板蓝)是其常用染材,而别名吴蓝、苋蓝则为此四类所涵括。植物虽有差别,却皆含有靛苷成分,故可以直接染色法、水煮染色法、发酵还原染色法等,将靛蓝附着在织物纤维上。千百年来,人们就地取材不断积累经验,更是在融入社会生活、习俗与政治、经济因素的同时,通过广泛地交流形成独具特色的蓝染文化圈。

辽东地区高羊茅和早熟禾公路护坡特性研究

中国公路迅猛发展,开发出的边坡需要种植相应的草种防止水土流失。通过对高羊茅和早熟禾的盖度、生长量、抗旱性、抗瘠薄性、土体崩解性、根系生物量和滞尘量来对草种的护坡特性进行试验分析。两个草种的盖度8-9月份分别达到最大值98%和99%,生长量在70~100 d达到最大值,抗旱性和抗瘠薄性均较强,根系总生物量分别为84.06 g和52.96 g,滞尘量和土体抗崩解性高羊茅优于早熟禾。两个草种盖度、生长量、根系生物量和根系生物量对水土保持起到了明显的作用,其抗旱性和抗瘠薄性使得草种能够更易栽植于边坡,滞尘特性能够明显改善周围空气质量。草本植物护坡具有明显的优势,边坡防护应尽可能选用两种草本植物护坡,单一草本植物护坡选用高羊茅更为适合。

Development of natural treatment system consisting of black soil and Kentucky bluegrass for the post-treatment of anaerobically digested strong wastewater

To develop a sound post-treatment process for anaerobically-digested strong wastewater,a novel natural treatment system comprising two units is put forward.The first unit,a trickling filter,provides for further reduction of biochemical oxygen demand and adjustable nitrification.The subsequent soil–plant unit aims at removing and recovering the nutrients nitrogen（N）,phosphorus（P）and potassium（K）.As a lab-scale feasibility study,a soil column test was conducted,in which black soil and valuable Kentucky bluegrass were integrated to treat artificial nutrient-enriched wastewater.After a long-term operation,the nitrification function was well established in the top layers,despite the need for an improved denitrification process prior to discharge.P and K were retained by the soil through distinct mechanisms.Since they either partially or totally remained in plant-available forms in the soil,indirect nutrient reuse could be achieved.As for Kentucky bluegrass,it displayed better growth status when receiving wastewater,with direct recovery of 8%,6% and 14% of input N,P and K,respectively.Furthermore,the indispensable role of Kentucky bluegrass for better treatment performance was proved,as it enhanced the cell-specific nitrification potential of the soil nitrifying microorganisms inhabiting the rhizosphere.After further upgrade,the proposed system is expected to become a new solution for strong wastewater pollution.

草地早熟禾蛋白激酶CIPK24基因克隆及非生物胁迫响应分析

CIPK蛋白家族是Ca^(2+)介导的植物信号通路中关键蛋白家族,当植物受到逆境胁迫时,CBL-CIPK网络可调节离子运输进而适应逆境环境,其中CIPK24作用显著。为探究禾本科草坪草CIPK家族基因的作用,本研究以草地早熟禾(Poa pratensis L.)为试验材料,采用RT-PCR技术对草地早熟禾CIPK24基因进行克隆,并应用生物信息学及qRT-PCR技术,对非生物胁迫条件下该基因的表达方式及组织特异性进行研究。研究结果显示,草地早熟禾CIPK24包含典型的结构域CIPK_C,其属于AMPKA_Clike superfamily,与黑麦草(Lolium perenne L.),大麦(Hordeum vulgare L.),二粒小麦(Triticum turgidum L.)CIPK24同源性高达94.20%,90.62%,88.62%。草地早熟禾CIPK24基因的表达水平存在组织特异性,叶>茎>根。CIPK24基因积极响应干旱、氮素、磷素、盐胁迫,其中,无氮、低氮、无磷、高盐环境显著促进其表达。本研究为丰富草地早熟禾CIPK家族在非生物胁迫下的调节机制提供了理论基础。

镉在草地早熟禾中的分布和化学形态研究

为探究草地早熟禾(Poa pratensis L.)对重金属镉(Cadmium,Cd)的分配和耐受机制,以草地早熟禾耐Cd品种‘午夜’(‘Midnight’,M)和Cd敏感品种‘橄榄球2号’(‘Rugby II’,R)为试验材料,在水培条件下施加600μmol·L^(-1)Cd胁迫,研究Cd在草地早熟禾中的组织分布、亚细胞分布及化学形态。结果表明,与R品种相比,Cd胁迫下M品种叶片失绿较少,生长抑制较小;表皮+叶肉、维管束及根表皮+皮层中的Cd含量显著低于R品种;另外,根系可溶部分Cd含量高于R品种。进一步分析发现,R品种含有更多毒性较高的无机态Cd及果胶和蛋白质结合态Cd,而M品种水溶性有机酸结合的Cd在根表皮+皮层、中柱等组织中含量较多,草酸盐结合的Cd在根系中柱含量显著高于R品种。因此,M品种通过减少Cd吸收、增加液泡区隔化、增加有机酸盐结合的Cd并降低Cd向地上的长距离运输来增强对Cd的抗性。

甘肃野生草地早熟禾种质种子产量相关性状分析及其对矿质元素利用效应评价

本研究以甘肃地区6份野生草地早熟禾种质为材料,开展了其种子产量相关性状特征分析及其对矿质元素利用效应评价,旨在为甘肃野生草地早熟禾种质资源利用改良和种质创新以及对矿物质高效利用提供研究基础和理论参考。结果表明,11个种子产量相关性状和8种矿质元素的变异系数为4.46%~50.88%,其中钠、钙、分枝上的种子数变异系数均大于40.00%,变异幅度较大。相关性分析结果显示,增加花序长、分枝数、小穗上的小花数、分枝上的种子数、小穗上的种子数以及小穗结实率,可提高草地早熟禾种子产量。8种矿质元素的平均含量顺序为K>Mg>Ca>Na>Mn>Zn>Fe>Cu,且不同元素之间存在复杂的相关性,以此为依据得出草地早熟禾生殖生长期间,多施K、Cu和Mn肥,少施Zn、Fe、Na、Mg和Ca肥可获得种子高产。聚类和主成分分析结果显示,依据19个性状对甘肃野生草地早熟禾的综合排名为秦州>兰州>陇西>甘南>清水>陇南,且前2个、中间2个及后2个种质各为一类,表明秦州这个材料在小穗上的小花数、分枝上的种子数、小穗上的种子数、花序种子数,小穗结实率、千粒重上均表现较好,可作为育种研究的优良亲本材料。

基于Web of Science与CNKI数据库的草地早熟禾文献计量分析

草地早熟禾是世界上优良的冷季型草坪草,具有色泽浓绿、绿期长、适应性强等特点。为了解目前国内外草地早熟禾的研究现状及发展方向,利用文献计量学方法,对Web of Science核心合集数据库(WOS)和中国知网数据库(CNKI)中草地早熟禾近20年的年度发文量、国家间合作关系、所属机构、文献来源及关键词等方面进行了梳理。结果表明:近20年来,两个数据库中收录的关于草地早熟禾文章数量随时间变化呈上升趋势。美国、中国、加拿大的发文量位于世界前列,各个国家合作紧密。美国罗格斯大学(Rutgers State University)和甘肃农业大学的发文量最多。国内外的期刊中《草业科学》和Crop Science的载文量最多。在以“Kentucky bluegrass/草地早熟禾”为核心的关键词共现网络图谱中,与逆境胁迫相关的关键词(“Resistance”“Response”“Tolerance”“Stress”“Drought”“抗旱性”“抗寒性”“干旱胁迫”“镉胁迫”等)出现频率较高。文章总结过去20年草地早熟禾的研究热点,将为从事草地早熟禾相关研究的科技工作者未来的科研方向提供指导。

外源IAA作用下草地早熟禾中调控Cd长距离运输的关键基因表达及其代谢通路分析

为探究吲哚乙酸(indole-3-acetic acid,IAA)对草地早熟禾中镉(cadmium,Cd)从根系到叶片长距离运输的影响及相关机制,分析IAA作用下草地早熟禾响应Cd胁迫的差异基因及代谢通路。以草地早熟禾为试验材料,在水培条件下,叶面喷施400 nmol·L^(-1)外源IAA并施加600μmol·L^(-1)Cd胁迫。进行株高、根长、根系和叶片Cd含量测定及转录组测序。利用实时荧光定量验证转录组数据准确性。结果表明,外源施加IAA可促进Cd胁迫下草地早熟禾株高和根长,抑制Cd从根系向叶片的长距离运输。转录组分析发现,Cd胁迫下共有1294条基因被IAA显著调控。实时荧光定量结果与RNA-Seq表达模式一致。GO富集分析表明,IAA可调控响应高温、低温、Cd离子及病毒的相关基因抵御Cd胁迫,其中分子伴侣蛋白HSP70、激酶MAPK和转录因子MYB46被明显上调。另外,外源IAA可上调谷胱甘肽代谢及木质素合成通路上的基因,可能增加谷胱甘肽合成及细胞壁木质化,进一步增加液泡区隔化和降低Cd进入细胞,从而减少叶片Cd含量。本研究可为IAA信号作用下Cd转运与分配机制研究提供参考。

30份草地早熟禾苗期耐盐性综合评价

为了发掘耐盐性强的优良草种质资源,本研究对常见的30份商用草地早熟禾(Poa pratensis)品种苗期的耐盐性进行鉴定评价。在400 mmol·L-1 NaCl胁迫下,测定了草地早熟禾的株高、分蘖数、生物量、叶绿素含量和叶片相对含水量等一系列生长和生理指标,并采用主成分分析法对草地早熟禾耐盐性进行综合评价。结果显示:在盐胁迫下,不同品种草地早熟禾的生长均受到抑制。其中‘肯塔基’的株高和地上部鲜重受盐胁迫的影响最小,与无盐对照相比,分别下降了32.62%和18.14%,而‘蓝之梦’地上部生物量受盐胁迫的影响最大,其地上部鲜重和干重分别较对照下降了71.80%和53.29%;与对照相比,盐处理后‘肯塔基’‘优美’‘月光’‘探险家’和‘乐土’的叶绿素含量下降幅度较低,均下降了20%上下,‘蓝精灵’叶绿素b含量下降高达71.40%,‘满月’的总叶绿素含量下降了55.70%。盐胁迫下,‘蓝精灵’‘满月’和‘优异’的叶片相对含水量下降幅度较大,超过了22%。结合主成分分析综合得分,耐盐性较强的草地早熟禾品种有‘肯塔基’‘乐土’‘探险家’‘月光’和‘优美’,而‘蓝精灵’‘飞达’‘优异’‘蓝之梦’和‘满月’对盐胁迫较为敏感。

5份草地早熟禾种子产量性状相关指标综合评价与分析

为了探究与草地早熟禾种子产量相关指标间的差异性以及不同指标对种子产量贡献率的大小。以不同来源的5份草地早熟禾为材料,测定16个种子产量相关指标,以期为筛选优良种质和新品种选育提供基础数据。结果显示,16个种子产量相关指标均呈正态分布,千粒重变异系数最小,改良潜力较小;每花序种子数变异系数最大,可选择范围较宽。相关性分析结果显示,每小穗种子数和每分枝种子数均与每花序结实率呈显著正相关,与千粒重呈显著负相关,表明种子数并不是越多越好,种子数越多,其质量越差。主成分和聚类分析结果显示,第Ⅰ主成分为每小穗种子数、每分枝种子数等产量因子,第Ⅱ主成分为花序长、分枝长等形态因子;依据16个指标综合排名最高的草地早熟禾种质为甘谷,且甘谷和定西为第Ⅰ类群,可作为种子高产育种材料,肃南和青海扁茎为第Ⅱ类群,可作为饲草资源利用,‘午夜2号’为第Ⅲ类群,可用于草坪建植及发展籽粒有关的副产品。

不同早熟禾品种对干旱胁迫的生理响应及抗旱性评价

为鉴定评价早熟禾幼苗抗旱性,将14个早熟禾品种的种子经消毒后播种在塑料花盆中,出苗后间苗,2-3个真叶定株,每盆留生长一致、分布均匀的幼苗15株,定期浇水,以保证幼苗的正常生长。待生长到3-4个真叶时进行干旱处理,依据Michel公式计算不同水势所需的PEG-6000的量,然后加入PEG-6000溶液进行干旱处理,处理的水势梯度为0MPa（即为对照）,-0.3,-0.6,-0.9和-1.2MPa。处理后每2d进行称重补水以维持溶液浓度。设干旱处理与对照（正常浇水）两组,对14个早熟禾品种幼苗进行模拟干旱胁迫,4次重复。测定叶绿素（CHL）含量、丙二醛（MDA）含量、脯氨酸（Pro）含量、过氧化物酶（POD）以及超氧化物歧化酶（SOD）活性等生理生化指标。结果表明,随着干旱胁迫的加剧,早熟禾幼苗叶片CHL含量呈下降趋势;Pro含量、MDA含量、SOD活性总体呈上升趋势;POD活性总体呈先降再升后降的趋势;利用隶属函数分析得出14个早熟禾品种的抗旱性强弱顺序依次为：3号恩托佩草地早熟禾〉10号细叶早熟禾〉1号优异草地早熟禾〉2号斗士草地早熟禾=9号蓝钻草地早熟禾〉7号草地早熟禾〉11号草地早熟禾〉4号草地早熟禾〉5号瓦巴斯草地早熟禾〉8号早熟禾=13号肯塔基草地早熟禾〉14号草地早熟禾〉6号莫诺波利草地早熟禾=12号菲尔津草地早熟禾。