青海草地早熟禾(Poa pratensis L.cv.Qinghai)近几年的研究进展

论述了青海草地早熟禾(Poa pratensis L.cv.Qinghai)野生栽培品种的由来,以及近几年在生物-生态性、恢复生态学、生理生化和高寒草坪方面的工作,提出了青海草地早熟禾目前依然缺乏的研究,指出今后工作的重点,应加强青藏高原育种工作,寻找更多更好的牧草生态型品种。

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.

冷地型草坪草耐热机理初探——Ⅰ.草地早熟禾(Poa pratensis L.)在热境胁迫下叶片叶绿素含量和POD酶活性的变化

草地早熟禾在持续高温（37～38℃）胁迫下表现为叶绿素相对含量下降,POD酶活性在处理2d时表现为下降,4d及4d后则明显提高。耐热品种较不耐热品种的叶片POD酶活性无论是适温（25℃）还是高温（38℃）下一般均较高。热境下有较高的POD酶活性可能是植株体耐热的主要原因之一。

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.

Genetic Relationships in Advanced Generation Hybrids Derived from Crosses between Texas (<i>Poa arachnifera</i>) and Kentucky (<i>Poa pratensis</i>) Bluegrass Using ISSR Markers

Fertile, advanced generation hybrids derived from crosses between Texas (Poa arachnifera Torr.) and Kentucky (Poa pratensis L.) bluegrass have been selected. The hybrids are currently being evaluated for low-input turf potential. Since they are derived from hand-harvested seed from first-generation hybrids located in field nurseries their exact genetic origin is unknown. This experiment was conducted to determine if there are still genetic relationships among the advanced generation hybrids and some of the Texas and Kentucky parents in their pedigrees. Four Inter Simple Sequence Repeat (ISSR) primer combinations resolved on 6% nondenaturing polyacrylamide gels resulted in 179 polymorphic bands that were scored to create a genetic similarity matrix and dendrogram based on Jaccard’s coefficient. The clustering of the advanced generation hybrids was generally in agreement with what would be expected based on their pedigrees and indicated it was more likely to select a fertile hybrid from an advanced generation, rather than the F1 generation.

Analysis of drought resistance HVA1 gene under drought stress in different Poa pratensis cultivars

Total RNA from leaves of Poapratensis cultivars under drought stress was extracted for reversing transcription to cDNA and then cDNA as template for PCR reaction by designing primer of cds of Hordeum valgare HVA1 drought resistance gene from GenBank. The amplified products were positive recon identified by using procedures of recovery, connection, transformation and enzyme separation. The length of cloned gene sequence was 324 bp, identity reached 79.27% with Barley HVA1 gene that meaned the cloned gene sequence was the partial HVA1 gene of Poapratensis.

The Complete Chloroplast Genome of <i>Poa pratensis</i>(Poaceae), a High-Quality Forage

Poa pratensis L. (Poaceae) is a hardy, persistent, attractive forage and turf grass adapted to a wide range of soils and climate. In this study, we release and detail the complete chloroplast genome sequences of P. pratensis. The whole chloroplast genome was 135,649 bp in length and comprised 131 genes, including 85 protein-coding genes, 38 tRNA genes, 8 rRNA genes. The P. pratensis chloroplast genome had a GC content of 38.3%. The result of phylogenetic analysis showed that P. pratensis was closely related to P. pratensis cv. Qinghai and P. poophagorum. This study would provide useful genetic information for the protection of P. pratensis and other related species.

Target-site Mechanisms Involved in Annual Bluegrass(Poa annua L.) Tolerance to Fenoxaprop-P-ethyl

Annual bluegrass （Poa annua L.） was found to be tolerant to fenoxaprop- P-ethyl as well as quizalofop-P-ethyl, haloxyfop-R-methyl, clodinafop-propargyl, fluaz- ifop-P-butyl, cyhalofop-butyl, sethoxydim and tralkoxydim, whereas it was sensitive to clethodim and tepraloxydim. The acetyI-CoA carboxylase （ACCase） IC50 values of five P. annua biotypes were 10.46 to 11.98-fold higher than the susceptible Japanese foxtail （Alopecurus japonicus Steud.）. The presence of the polymorphic lie and Leu at 1 781, which the presence of Leu at 1 781 had been reported to be in- volved in the resistance of grass weeds to ACCase inhibitors, was subsequently i- dentified in the ACCase of P. annua. Furthermore, the expression level of gene that encoding P. annua ACCase was found to be approximately 4.67 to 7.37-fold higher than A. japonicus, possibly explaining the P. annua target site tolerance to fenoxaprop-P-ethyl.

Effects of traffic pollution on the genetic structure of Poa annua L.populations

The genetic composition of Poa annua L. populations with a series of traffic pollution was studied by starch electrophoresis. Five enzyme systems were stained. The results showed that: (1) Traffic pollution can dramatically change genotypic frequencies at some loci of P. annua populations. Significant deviations from Hardy Weinberg equilibrium were observed on loci Fe 1 and Me due to the excess of heterozygotes in some populations. (2) The effective number of alleles per locus and the observed and expected heterozygosity were higher in the pollution series than in the clear control site(Botanic Park population), but the increase was not related with the pollution extent. (3) Most genetic variation was found within populations, and only 6 21% was among populations of the polluted series. Slightly higher differentiation( F ST =7 98%) was observed when the control population was included. (4) The calculated gene flow(Nm) is 2 8841 per generation. The mean of genetic identity is 0 9864 and the genetic distance average to 0 0138

Effects of Different Herbicides on the Control of Malachium aquaticum L. and Poa annua L.

In this experiment,the effects of 13 kinds of herbicides( or mixtures) on the control of two kinds of weeds were studied by pot cultivation with the broadleaf weed Malachium aquaticum L. and Gramineae weed Poa annua L. as the materials. The results showed that the herbicide MCPA-Na had the best and fastest control effect on M. aquaticum L.,and all the M. aquaticum L. died 7 d after treatment;it was followed by the other four herbicides including MCPA-Na + clethodim,MCPA-Na + quizalofop-p-ethyl,bentazon and nicosulfuron·atrazine,and all the M. aquaticum L. died 14 d after treatment. Atrazine was the best herbicide to control Gramineae weeds,followed by nicosulfuron·atrazine,and mesotrione·nicosulfuron·atrazine. The study on the application of field herbicide found that four herbicides including atrazine,mesotrione·nicosulfuron·atrazine,nicosulfuron·atrazine and bentazon had better control effect on weeds. The best herbicide for flax field was MCPA-Na + clethodim,followed by MCPA-Na and MCPA-Na + quizalofop-p-ethyl. The optimized herbicides and combinations had no harmful effects on the growth of corn and flax.

Distribution Area and Yield Indicators of Poa bulbosa L. in Uzbekistan

The article is based on field research, existing literature, and samples of herbarium kept in the National Herbarium fund (TASH) Poa bulbosa L. The distribution maps and productivity indicators in the flora of Uzbekistan are presented. As you know, Poa bulbosa L. is considered the wild relative of cultural plants, as a forage plant, it is a plant with high protein value. In the course of research during 2021-2022, in 16 areas, the yield indicators of the species were determined. In the studied research areas, indicators of the total weight of plant bushes (on the account of wet mass) and, respectively, the volume of plant biomass (on the account of dry mass, the upper part of 25%) were determined. According to him, it was determined that the average yield of the plant the wet mass is 14669.8 kg/ha, and the average yield of the plant the dry mass in areas of total 244.0 ha in the total area is 63172.5 kg.

Agrobacterium-mediated Transformation of Kentucky Bluegrass

Embryogenic calli of Kentucky bluegrass, named Md, were induced from mature seeds and embryos, and proliferated on medium K3 containing 2,4-dichlorophenoxyacetic acid (2,4-D, 10.0 mumol/L), 6-benzylaminopurine (BAR, 0.5 mumol/L) and K5 which was the K3 medium supplemented with cupric sulfa (0.5 mumol/L) under dim-light condition (20-30 mumol.m(-2).s-1, 16 h light) at 24 degreesC. Embryogenic calli were transformed with plasmids pDM805 Carring bar and gus genes, Which was mediated by an Agrobacterium strain AGL1, four transgenic lines were obtained. The important factors that affect the transformation efficiency and obtain desirable number of transgenic plants included: (1) the quality of embryogenic calli; (2) light condition and time of co-cultivation; (3) concentration of antibiotics used for suppressing the overgrowth of Agrobacterium in the course of transformed plant regeneration; (4) selection pressure, etc. The micro nutrient of cupric had significant influence on the quality of embryogenic calli. This presentation is the first successful protocol of Kentucky bluegrass transformation mediated by Agrobacterium.

草地早熟禾叶绿素酶1基因PpCHL1的克隆和表达分析

利用RACE技术首次从草地早熟禾中克隆出叶绿素酶1基因(PpCHL1)的全长cDNA序列,提交到GenBank,登录号为KU145126;其开放阅读框为951bp,编码361个氨基酸,等电点6.11,平均亲水指数0.084,属于疏水性酸性蛋白。高级结构分析表明,其具有脂肪酶家族的保守域及水解酶特异的功能活性位点。利用实时荧光定量PCR分析其在草地早熟禾根、茎和叶及三种植物生长调节剂脱落酸(ABA)、水杨酸(SA)和茉莉酸甲酯(MeJA)处理后的表达情况,结果显示PpCHL1基因在草地早熟禾不同组织的表达情况存在明显的组织差异性,叶中的表达量最高且受ABA、SA和MeJA诱导。

来源于海洋真菌的Oxalicumone A诱导肝细胞L-02凋亡的毒性作用机制研究

目的评价来源于海洋真菌Penicillium sp.的新药候选化合物Oxalicumone A(POA)对人正常肝细胞L-02的细胞毒性作用,初步探讨POA诱导其凋亡的作用机制。方法以体外培养的L-02为实验对象,CCK-8检测细胞存活率;Hoechst 33258染色观察细胞形态学的改变;流式细胞仪检测细胞凋亡率和增殖周期;Western blot检测凋亡相关蛋白Fas、Bax以及Bcl-2的表达。结果 CCK-8检测结果表明,POA对L-02细胞有一定的时间、浓度依赖性的抑制作用;Hoechst 33258染色可见细胞出现核固缩、碎裂、浓染等典型的凋亡细胞形态学改变;流式细胞仪检测显示细胞早期凋亡率和Sub-G1、S、G2/M期出现了相应比例的增加;Western blot法表明POA显著增加了凋亡蛋白Fas、Bax表达(P<0.05),降低了抗凋亡蛋白Bcl-2表达(P<0.05)。结论 POA体外诱导L-02细胞毒活性,产生凋亡并经过线粒体信号通路,其作用机制与Bcl-2表达下调以及Fas、Bax表达上调有关。

Effects of Drought Stress on Photosynthetic Characteristics of Kentucky Bluegrass

Drought stress is one of the major constraints on turf growth and performance in northern China. Based on previous assessments of drought tolerance levels, four varieties of Kentucky bluegrass （Poa pratensis L.） representing different types （＇Baron＇, BVMG type; ＇Blackstone＇, Cheri type; ＇Kenblue＇, Midwest type; and ＇Bluemoon＇, Midnight type） were chosen for this study. Grass materials were established in a glasshouse, and the changes in their photosynthetic and metabolic characteristics were analyzed during a period of imposed drought stress. Drought stress led to decrease in chlorophyll content, photosynthetic rate, stomatal conductance and transpiration rate in all four cultivars, but the relative changes in each characteristic varied among varieties. The results also showed that changes in the photosynthetic and metabolic characteristics of different varieties under drought stress were related to the morphological characteristics of the plants. Varieties with longer narrow leaves, longer roots, higher root-to-shoot ratios, and higher chlorophyll contents showed greater rates of photosynthesis and stomatal conductance, and lower transpiration rates.

NaCl胁迫对3种草地早熟禾种子萌发特性的影响

用室内模拟的方法,对"午夜""新歌来德""抢手股"3种草地早熟禾种子进行4种不同浓度(0、1、1.5、2mmol·L-1)的NaCl溶液盐胁迫试验,结果表明:盐浓度越高,植物受到的盐胁迫越大;3种早熟禾种子中,发芽率、胚根长度和胚芽长度最高的均为午夜种子,最低的为抢手股种子。午夜种子的综合能力较强,最适合应用于抗盐碱土地中,新歌来德次之,而抢手股不适合种植在盐碱地中。

硝酸钾配施有机肥对早熟禾生长及养分吸收的影响

早熟禾草坪在盐碱土壤上建植3~4年后会发生明显的退化,研究园林养护中采用硝酸钾配施有机肥的方式为其提供营养,对促进植株生长和提高坪用质量具有重要意义。试验设置在城市绿化带中,设置空白对照(K1)、硝酸钾化肥对照(K2)、化肥+发酵饼肥(K3)、化肥+腐殖酸复混肥(K4)4个处理,每个处理3次重复,研究了早熟禾生长和养分吸收变化。结果表明:K3与K1相比,在7~10月有利于提高早熟禾干物质积累量、含水量、叶绿素a、叶绿素b和类胡萝卜素含量,显著提高了植株氮、磷、钾吸收量;K3与K2相比,在8~9月显著提高了干物质积累量,7月和10月显著提高了叶绿素b和类胡萝卜素含量,8~10月提高了氮磷钾养分吸收量;K3与K4相比,在7~9月提高了干物质积累量,7~10月显著提高了叶绿素b和类胡萝卜素含量,促进了氮磷钾营养元素的吸收。综合分析认为,硝酸钾配施发酵饼肥为适宜施肥处理。

硝酸钾配施发酵饼肥对景观草坪坪用质量的影响

通过研究硝酸钾配施发酵饼肥对草地早熟禾草坪坪用质量影响的规律,可以为城市园林草坪养护的科学施肥提供理论依据。试验在城市环境条件下,设置对照(CK)、硝酸钾(K_(1))、硝酸钾+发酵饼肥(K_(2))、硝酸钾+腐殖酸复混肥(K_(3))4个处理,小区试验设计,3次重复。结果表明:6月,K_(1)处理分蘖密度、株高、叶长、叶绿素含量均显著高于CK、K_(2)和K_(3)处理;7-10月,K_(2)分蘖密度显著高于CK、K_(1)和K_(3),6-8月K_(1)与K_(3)均显著高于CK;6-10月,K_(2)处理盖度显著高于CK,6-7月和9-10月K_(2)显著高于K_(1);7-10月K_(2)株高和叶长均显著高于CK,8月和10月K_(2)显著高于K_(1);7月、8月和10月,K_(2)处理叶宽显著高于对照;7-10月,K_(2)处理叶绿素含量和植株鲜重均显著高于CK,K_(2)显著高于K_(3)。综合分析认为,在现有施肥量基础上硝酸钾与发酵饼肥配合施用有利于显著提高草地早熟禾草坪坪用质量。

草地早熟禾苗期耐低温的生理特性研究

为了探究不同抗性草地早熟禾苗期对低温胁迫的生理特性差异,以10个常用草地早熟禾品种为试验材料,将幼苗叶片进行不同程度的低温胁迫处理,利用细胞伤害率和对应的低温度梯度进行Logistic方程拟合,获得半致死温度(LT50)并筛选抗寒品种和低温敏感品种,并以二者为试验材料进行相关生理指标的测定和相关性分析。结果表明,草地早熟禾‘普瑞芬’的LT_(50)为-17.5℃,表现出较好的抗寒性,‘帝国’的LT_(50)为-3.2℃,表现出对低温的敏感性;经低温胁迫处理后抗寒品种‘普瑞芬’的相对电导率和丙二醛含量显著下降并低于低温敏感品种‘帝国’,而‘普瑞芬’幼苗中干物质含量、相对含水量和苗长等生长指标和叶绿素含量、渗透调节物质(游离脯氨酸、可溶性糖和可溶性蛋白)和抗氧化酶活性(SOD、POD和CAT)均高于‘帝国’;经相关性分析表明,低温对草地早熟禾幼苗的干物质,苗长和根长及渗透调节物质含量影响较大且各指标之间相互作用。

草地早熟禾PpSWEET1b基因克隆及功能研究

SWEETs糖转运蛋白是一类在真核生物和原核生物中均存在的促进糖通过细胞膜流动的转运蛋白之一,参与调控植物的生长发育和胁迫响应过程。以草地早熟禾品种蓝月为材料,以叶片cDNA为模板克隆PpSWEET1b基因,采用花序侵染法转化拟南芥,获得了PpSWEET1b拟南芥过表达株系,并研究干旱和低温胁迫对转基因和野生型拟南芥生长、生理及基因表达的影响。结果表明:干旱和低温胁迫下,PpSWEET1b过表达植株生长状态均优于野生型,叶片MDA含量显著低于野生型,但葡萄糖含量较野生型显著增加,抗旱性和抗寒性明显增强;PpSWEET1b过表达植株可能协同SWEET2和SWEET3共同参与了对低温的响应。此外,以叶片DNA为模板克隆了PpSWEET1b基因的启动子序列,顺式作用元件预测发现,其含有多个与胁迫和激素响应、光响应相关的元件。综合以上结果表明,草地早熟禾PpSWEET1b基因在耐旱和耐寒方面发挥着重要作用,可为草坪草抵抗非生物胁迫生物育种提供新的优异基因资源。