非结构糖在柠条(Caragana korshinskii K.)再生生长和持续生存中的作用

柠条在地上组织破坏后从根颈处萌蘖出大量的一年生枝,根系储存的碳水化合物将供应地上生长。但连续刈割后根系储存物质如何变化,储存物质在柠条生长和维持存活中的作用仍须研究。本文在不同时间段内对柠条新萌蘖枝进行连续去除(处理1:对照,3月28日刈割1次后不再处理;处理2:从萌芽到6月9日,每3 d去除一次新萌发枝;处理3:从萌芽到6月24日,每3 d去除一次新萌发枝;处理4:从萌芽到7月15,每3 d去除一次新萌发枝)。结果表明:处理3和4在7月非结构糖含量显著高于处理1和2,说明处理1和2将根系储存的非结构糖用于地上生物量的恢复。连续刈割处理(处理3和4)导致柠条根系非结构糖含量持续下降,这主要是由根系呼吸作用引起。秋末对当年生枝统计结果表明,各处理之间当年生枝数/刈割枝在各处理间差异不显著,说明柠条对外界的破坏具有极强的忍耐能力。

Spatiotemporal characteristics of seed rain and soil seed bank of artificial Caragana korshinskii Kom. forest in the Tengger Desert, China

Vegetation restoration and reconstruction are effective approaches to desertification control and achieving social and economic sustainability in desert areas.However,the self-succession ability of native plants during the later periods of vegetation restoration remains unclear.Therefore,this study was conducted to bridge the knowledge gap by investigating the regeneration dynamics of artificial forest under natural conditions.The information of seed rain and soil seed bank was collected and quantified from an artificial Caragana korshinskii Kom.forest in the Tengger Desert,China.The germination tests were conducted in a laboratory setting.The analysis of species quantity and diversity in seed rain and soil seed bank was conducted to assess the impact of different durations of sand fixation(60,40,and 20 a)on the progress of vegetation restoration and ecological conditions in artificial C.korshinskii forest.The results showed that the top three dominant plant species in seed rain were Echinops gmelinii Turcz.,Eragrostis minor Host.,and Agropyron mongolicum Keng.,and the top three dominant plant species in soil seed bank were E.minor,Chloris virgata Sw.,and E.gmelinii.As restoration period increased,the density of seed rain and soil seed bank increased first and then decreased.While for species richness,as restoration period increased,it gradually increased in seed rain but decreased in soil seed bank.There was a positive correlation between seed rain density and soil seed bank density among all the three restoration periods.The species similarity between seed rain or soil seed bank and aboveground vegetation decreased with the extension of restoration period.The shape of the seeds,specifically those with external appendages such as spines and crown hair,clearly had an effect on their dispersal,then resulting in lower seed density in soil seed bank.In addition,precipitation was a crucial factor in promoting rapid germination,also resulting in lower seed density in soil seed bank.Our findings provide valuable insights for guiding future interventions during the later periods of artificial C.korshinskii forest,such as sowing and restoration efforts using unmanned aerial vehicles.

水分胁迫下AM真菌对柠条锦鸡儿(Caragana korshinskii)生长和抗旱性的影响

利用盆栽实验研究了水分胁迫条件下AM真菌对柠条锦鸡儿（Caragana korshinskii）生长和抗旱性的影响。在土壤相对含水量为80%、60%和40%条件下,分别接种摩西球囊霉（Glomus mosseae）和柠条锦鸡儿根际土著菌,结果表明,水分胁迫对AM真菌的接种效果有显著影响。不同水分条件下,接种AM真菌显著提高了宿主植物根系菌根侵染率。土壤相对含水量为40%-60%时,接种株的株高、茎粗、生物干重和叶片保水力明显高于不接种株;接种AM真菌提高了植株对土壤有效N和有效P的利用率,增加了植株全P、叶片叶绿素和可溶性糖含量以及SOD、POD、CAT等保护酶活性。土壤相对含水量为40%时,叶片MDA含量明显下降。水分胁迫条件下,以接种柠条锦鸡儿根际土著菌的效果最佳。AM真菌增强宿主植物的抗旱性可能源于促进宿主植物根系对土壤水分和矿质元素吸收的直接作用和改善植物体内生理代谢活动、提高保护酶活性的间接作用。

柠条（Caragana korshinskii）的水分生理生态学特性

柠条的水分生理生态学研究表明．蒸腾速率为３９５．１±２３４．４ｍｇ·ｇ－１·ｈ－１（平均值±标准差）；小枝水势为－１．５８８ＭＰａ；叶含水率为６０．１２±３．２９％；蒸腾系数为７２．８２±１９．８８ｇＨ２Ｏ·ｇＤＷ－１。林地土壤水分含量与蒸腾速率、小枝水势和叶含水率的关系十分密切。响应于年生长发育节律和水分条件的变化，柠条的抗旱性在年生长期初（５月）和旺盛生长阶段（８～９月）较弱．在高温干旱阶段（６～７月）和入冬之前（１０月）较强。在年生长期中间（６～９月），柠条的抗旱性与蒸腾速率、小枝水势和叶含水率之间均具有负相关性。

Interspecific Transition Among Caragana microphylla, C. davazamcii and C. korshinskii Along Geographic Gradient. Ⅰ. Ecological and RAPD Evidence

The former plant population survey has shown that three genetically-related species, Caraganamicrophylla Lam., C. davazamcii Sancz. and C. korshinskii Kom., form a geographical replacement series inNei Mongol Plateau. The present study on population distribution, taxonomy, morphology, development andgenetic structure demonstrated that the geographical distribution of these three species was successiveand in gradual change, thus forming a geographical cline which extended from the east to the west of NeiMongol Plateau. With an analysis of climate change over time, it was considered that the formation of thisgeographical cline was a result of plant adaptation to its natural environment.

Interspecific Transition Among Caragana microphylla, C. davazamcii and C. korshinskii Along Geographic Gradient. Ⅱ. Characteristics of Photosynthesis and Water Metabolism

The characteristics of photosynthesis and water metabolism of Caragana microphylla Lam.,C. davazamcii Sancz. and C. korshinskii Kom. populations in different sites (117.6o-105.7o E, 44.6o-38.8o N)were studied. (1) From the east to the west, the responses of the three species to photosyntheticallyavailable radiation (PAR) in net photosynthesis rate increased, the relative humidity of the air whichcorresponded to the occurrence of maximum photosynthesis rate decreased, and the corresponding airtemperature increased. Along the same gradient, the before-noon superiority of the photosynthesis be-came evident, and the photosynthesis rate and the light use efficiency (LUE ) increased, while the transp-iration rate decreased, thus the water use efficiency (WUE ) increased notably, and the leaf water contentdecreased gradually. From the east to the west, the plants took a water-saving strategy step by step withhigher photosynthesis rate and lower transpiration rate. These physiological changes in the plants wereadaptable to the conditions of light, temperature and humidity in the habitat of the plants, and might be thebiological foundation for the geographical transition among C. microphylla , C. davazamcii and C. korshinskii.(2) The adaptation of photosynthetic system of C. microphylla , C. davazamcii and C. korshinskii to PAR, airhumidity and temperature exhibited the interspecific continuity, which was consistent with theenvironmental gradient. In different species and different sites, the diurnal changes of net photosynthesisrate, the daily cumulative value of net photosynthesis, the diurnal changes of transpiration rate, the dailycumulative value of transpiration, the water use efficiency and the diurnal changes of leaf water contentvaried with longitudinal descent (from the east to the west). The characteristics of photosynthesis andwater metabolism indicated that the geographical transition among C. microphylla , C. davazamcii and C.korshinskii was in gradual change, and these three species formed a geographical cline.

内蒙古高原小叶锦鸡儿(Caragana microphylla)中间锦鸡儿(C．davazamcii)和柠条锦鸡儿(C．korshinskii)的遗传多样性及遗传关系

采用RAPD技术对10个具较大地理跨度的小叶锦鸡儿、中间锦鸡儿和柠条锦鸡儿种群的遗传多样性和遗传关系进行了研究。共检测到678个位点,多态条带比率(PPB)为100%;特有位点41个,占6.05%。总体上3种锦鸡儿的遗传多样性表现出自东向西递减的趋势,分析表明其与生长地点年均气温呈显著负相关。AMOVA表明:3种锦鸡儿种间变异只占总体变异的6.08%,且显著性检验表明这种变异不显著;种内种群间的变异占总变异的11.90%;总变异的主要部分来自种群内部(82.02%)。3种锦鸡儿各种群总体分析结果表明:种群内变异比率Hpop/Hsp为0.8013,基因分化系数Gst为0.1603,种群每代迁移数Nm为2.6192,显示种群间存在一定强度的基因流,3种锦鸡儿间表现为异交性。3种锦鸡儿多样性高低及种群聚类分布格局都表现出一定的地理连续性。

Characteristics of root pullout resistance of Caragana korshinskii Kom.in the loess area of northeastern Qinghai-Tibet Plateau,China

Roots exert pullout resistance under pullout force,allowing plants to resist uprooting.However,the pullout resistance characteristics of taproot-type shrub species of different ages remain unclear.In this study,in order to improve our knowledge of pullout resistance characteristics of taproot systems of shrub species,we selected the shrub species Caragana korshinskii Kom.in different growth periods as the research plant and conducted in situ root pullout test.The relationships among the maximum pullout resistance,peak root displacement,shrub growth period,and aboveground growth indices(plant height and plant crown breadth)were analyzed,as well as the mechanical process of uprooting.Pullout resistance of 4-15 year-old C.korshinskii ranged from 2.49(±0.25)to 14.71(±4.96)kN,and the peak displacement ranged from 11.77(±8.61)to 26.50(±16.09)cm.The maximum pullout resistance and the peak displacement of roots increased as a power function(R^(2)=0.9038)and a linear function(R^(2)=0.8242)with increasing age,respectively.The maximum pullout resistance and the peak displacement increased with increasing plant height;however,this relationship was not significant.The maximum pullout resistance increased exponentially(R^(2)=0.5522)as the crown breadth increased.There was no significant relationship between the peak displacement and crown breadth.The pullout resistance and displacement curve were divided into three stages:the initial nonlinear growth,linear growth,and nonlinear stages.Two modes of failure of a single root occurred when the roots were subjected to vertical loading forces:the synchronous breakage mode and the periderm preferential breakage mode.These findings provide a foundation for further investigation of the soil reinforcement and slope protection mechanisms of this shrub species in the loess area of northeastern Qinghai-Tibet Plateau,China.

Succession of soil bacterial and fungal communities of Caragana korshinskii plantation in a typical agro-pastoral ecotone in northern China over a 50-a period

Bacterial and fungal communities play critical roles in reestablishing vegetation structure,function and biodiversity in ecosystem restoration in arid and semi-arid areas.However,the long-term successional changes in bacterial and fungal communities that occur with artificial vegetation development are not fully understood.In this study,we investigated the successional changes in bacterial and fungal communities in Caragana korshinskii Kom.plantation over a period of 50 a(6,12,18,40 and 50 a)and their relationships with key soil environmental factors in a typical agro-pastoral ecotone,northern China.The results showed that bacterial and fungal diversities(α-andβ-diversity)were significantly affected by plantation age;moreover,the change in fungal community was more evident than that in bacterial community.Soil samples from 12 a plantation had the highest(P<0.05)bacterial and fungalα-diversity(i.e.,abundance-based coverage estimator(ACE)and Chao1 index)at 0-10 cm depth compared with other samples.However,soil samples from plantation at the late recovery stage(40-50 a)had the highestα-diversity at 10-20 cm depth.Soil bacterial community was not significantly affected by plantation age at the genus level;but,soil fungal community was significantly affected at the genus level.Overall,Mortierella and Chaetomium were the dominant genera at natural recovery stage(0 a);Inocybe was the dominant genus at the early recovery stage(6-12 a);Inocybe and Mortierella were the dominant genera at the mid-recovery stage(12-40 a);And Mortierella,Cladosporium and Humicola were the dominant genera at the late recovery stage(40-50 a).Redundancy analysis(RDA)showed thatβ-glucosidase activity,total nitrogen and soil organic carbon were closely associated with bacterial community composition,while alkaline phosphatase,urease activity and total nitrogen were associated with fungal community composition,indicating that changes in enzyme activity and soil nutrients were the most important determinants of dominant genera.Furthermore,pathogenic microorganisms(Cladosporium and Humicola)were dominant in soils from 40-50 a plantation,which may affect plant growth,resulting in the decline of C.korshinskii plantation.Overall,the findings of this study improve the understanding of ecological patterns of bacterial and fungal communities in artificial vegetation and provide an important scientific basis for comprehensive ecological restoration management in arid and semi-arid areas.

Eco-physiological mechanisms of Caragana korshinskii Kom. adaptation to extreme drought stress: Leaf abscission and maintaining stem chloroplast integrity

In order to study the eco-physiological mechanisms of C. korshinskii adaptation to extreme drought stress, we investigated the variations of water content in soil, leaves, and stems, the chlorophyll a and b and the carotenoid content in leaves and stems, as well as changes of chloroplast ultrastructure in 2-year-old C. korshinskii specimens during a progressive soil drought process （by ceasing watering until all leaves were shed） and a subsequent rehydmtion process. During the dehydration process, the chlorophyll a and b and carotenoid contents in the leaves decreased, as did the carotenoid content in the stems. During the 4-day rehydration process, the chlorophyll a and b and carotenoid contents in the leaves and stems increased and gradually returned to normal levels. During ongoing drought stress, chloroplasts in the leaves broke away from cell walls and appeared in the center of cells. Under severe drought stress, the mesophyll ultrastmcture and chloroplast configuration in leaves were irreversibly disturbed, as mani- fested by the inner and outer membranes being destroyed; the thylakoid system disintegrated, the starch grain disappeared, and parts of cell tissue were dismantled into debris. However, the mesophyll ultrasWacture and chloroplast configuration in the stems remained complete. This indicates that C. korshinskii utilizes leaf abscission to reduce the surface area to avoid damage from ex- treme drought stress, and maintains chloroplast integrity and a considerable amount of chlorophyll to enable a rapid recovery of photosynthesis under the rehydration process.

Experimental Study on Friction Characteristics of Caragana korshinskii Kom

Caragana korshinskii Kom, which is a kind of excellent shrubs, has strong resistance to windstorms, and it is also a kind of forage that is of high nutritional value as well as a forming fuel conversion resource that is of high caloric. Caragana korshinskii Kom is of high lignifications after growing for a few years and the toughness of it is considerably high. Currently in China, equipments of harvesting and processing for ripe crops can hardly finish the mechanized production for Caragana korshinskii Kom. Friction characteristics of woody material for Caragana korshinskii Kom under different conditions should be given when the relevant operation machinery is designed, which can provide physical parameters for transportation of Caragana korshinskii Kom as well as the designing of relevant machinery. The paper bases on the research of friction characteristics between Caragana korshinskii Kom whose diameter of 7, 10, and 13 mm under sampling directions of 0°, 45°,and 90° and machinery materials that are commonly used such as steel plate, rubber sheet and so on, and meanwhile the test considers factors such as different conditions of Caragana korshinskii Kom, different materials of machines, different angles and so on. The data strongly suggests that the average static, dynamic friction coefficient between Caragana korshinskii Kom and steel plate is 0.399 711 and0.353 022, respectively; The average static and dynamic friction coefficient between Caragana korshinskii Kom and rubber sheet is 0.965 178 and 0.883 667, respectively. The maximum of static and dynamic friction coefficients is when the angle between the direction of sampling and the direction of movement is vertical. As the angle increased, the dynamic and static friction coefficient decreased. The friction coefficient between Caragana korshinskii Kom and steel plate were increased with the increase of the diameter of Caragana korshinskii Kom, but the diameter have no effect on the dynamic friction coefficient between Caragana korshinskii Kom and rubber plate. The conclusion of this paper can provide references for research on machines that are relevant to transportation and particle forming of Caragana korshinskii Kom.

Effect of Caragana korshinskii Silage at Different Growth Stages on the Feeding of Livestock

[Objectives] Caragana korshinskii resources are rich in Ordos with a large planting area, but they have not been fully and reasonably utilized. This study aims to effectively convert C. korshinskii into high-quality feed, improve the quality of milk and meat of herbivores and increase production. [Methods] The utilization of C. korshinskii in feed was studied in various aspects, the factors were selected, the level was determined, and the multi-factor experiment—orthogonal experiment was conducted. [Results] The effect on the growth of sheep was in the order of silage > C. korshinskii feed > corn stalk silage > hay meal, and the effect on the growth of goats was in the order of hay meal > C. korshinskii feed > silage > corn stalk silage. [Conclusions] From the analysis of the range results, the best level combination was selected: in different growing seasons, C. korshinskii branch and leaf silage in full-bloom stage had the greatest effect on the growth of sheep and goats, and the fattening effect was the best;among the different silage methods of C. korshinskii, the C. korshinskii—corn stalk mixture silage had a good feeding effect for sheep and goats.

黄土高原半干旱区柠条(Caragana korsh inskii)树干液流动态及其影响因子

应用热脉冲技术在黄土高原神木县六道沟小流域于2006年6月13至25日测定了两种不同密度柠条(Caragana korshinskii)群落的树干液流动态。同时测量了土壤水分、太阳辐射、大气温度、相对湿度、风速、水汽压亏缺和作物参考蒸散等环境因子,并根据植物蒸腾的P-M公式,反推计算冠层导度。结果表明,除风速外,柠条树木液流与太阳辐射、大气温度、相对湿度、水汽压亏缺、作物参考蒸散均显著相关,且可用太阳辐射的线性表达式来估测。不同密度群落的日蒸腾量随叶面积指数增大而增加,叶面积指数为2.3的群落平均日蒸腾为3.83mm d-1m-2,而叶面积指数为1.1的林分平均日蒸腾1.64mm d-1m-2。冠层导度与气象因子关系复杂,当土壤水分不存在亏缺时,冠层导度与太阳辐射、大气温度、作物参考蒸散因子显著相关,与水汽亏缺和相对湿度因子无相关性;当土壤水分存在亏缺时,冠层导度与太阳辐射、大气温度、作物参考蒸散因子无相关关系,而与水汽亏缺和相对湿度因子显著相关。

干旱胁迫下柠条锦鸡儿叶片SSH文库构建及CkWRKY1基因克隆

为研究柠条锦鸡儿抗旱的分子机制,挖掘干旱响应相关基因,成功构建干旱胁迫下柠条锦鸡儿叶片的抑制性差减杂交(SSH)文库,获得1286条高质量的EST序列,平均长度475bp,拼接得到Unigenes645条。对这些Unigenes进行Blast比对注释后发现很多抗旱相关基因,例如LEA蛋白、DHN蛋白编码基因,MYB、bZIP、WRKY、NAC和bHLH转录因子等。通过RACE技术克隆其中1个WRKY转录因子的cDNA全长,命名为CkWRKY1,GenBank登录号为JX987095。CkWRKY1编码330个氨基酸的蛋白质,氨基酸序列中有1个"WRKYGQK"的WRKY转录因子家族保守序列,属于第Ⅱ类WRKY转录因子。利用实时荧光定量PCR技术检测发现,CkWRKY1基因在干旱处理后mRNA的表达水平明显上升,预示该转录因子可能与柠条锦鸡儿抗旱的分子机制相关。

柠条锦鸡儿CkNCED1基因启动子的克隆及表达分析

利用染色体步移(Genome walking)技术,克隆柠条锦鸡儿CkNCED1基因上游启动子序列。经顺式元件预测分析,该序列除基本的启动元件之外,还含有2个脱落酸诱导响应元件ABRE、此外还含有多个逆境相关的元件。将CkNCED1基因启动子区连接到pGWB533植物表达载体上,构建Promoter::GUS载体,GUS组织化学染色结果表明,CkNCED1基因在植物的叶、根、茎、花和角果的维管组织中均有表达,且在叶片中表达强度最高。以上研究确定了柠条锦鸡儿CkNCED1基因的表达部位和强度。进一步说明CkNCED1基因在ABA合成调控中发挥重要作用,为深入研究基因功能奠定基础。

柠条锦鸡儿CkLEA4基因克隆及表达分析

LEA蛋白是一种与植物抗逆相关的胚胎发育晚期丰富蛋白。该研究从已构建的柠条锦鸡儿干旱胁迫抑制削减杂交文库中筛选到1条LEA蛋白编码基因的部分序列,用RACE技术扩增得到该基因cDNA全长并对其进行了克隆。测序表明该基因cDNA长870bp,其中开放阅读框长510bp,编码169个氨基酸,推测蛋白分子量为17.03kD,等电点为9.3,是一种亲水蛋白。序列比对和系统进化分析表明,该基因属于LEA4基因家族成员,命名为CkLEA4。实时荧光定量PCR检测发现,CkLEA4基因在干旱、ABA和NaCl处理下均受到不同程度的诱导,说明CkLEA4基因可能与柠条锦鸡儿响应逆境胁迫有关。

转CkLEA4基因拟南芥种子萌发期的抗逆性分析

该研究在实验室前期研究的基础上,将受脱水、盐胁迫和ABA诱导的柠条锦鸡儿CkLEA4基因转入野生型拟南芥,并利用实时荧光定量PCR从8株纯合体中筛选出3个表达量不同的株系,比较野生型和转CkLEA4基因过表达拟南芥种子在不同胁迫处理下的萌发率,以探讨CkLEA4基因在植物抵抗逆境胁迫中的功能。结果发现:(1)在不同浓度NaCl、甘露醇及ABA处理下,转CkLEA4基因过表达拟南芥种子的萌发率均高于野生型,随着NaCl、甘露醇及ABA浓度增加,各株系萌发率均降低,但野生型的萌发率下降幅度均高于3个过表达株系,并且在200mmol/L NaCl和400mmol/L甘露醇处理下,过表达株系子叶绿化率均显著高于野生型。(2)在低浓度ABA处理下,CkLEA4过表达植株子叶的绿化率也高于野生型。研究表明,柠条锦鸡儿CkLEA4基因提高了拟南芥种子萌发阶段对盐、ABA及渗透胁迫的耐受性。

柠条锦鸡儿CkCDPK基因的克隆、表达分析及载体构建

钙依赖蛋白激酶作为一个关键的信号传导器,通过调控和参与植物体内的代谢途径等方式在植物抵御逆境胁迫过程中发挥着重要作用。为了探索柠条锦鸡儿相关基因的功能,本研究利用同源克隆的方法,从柠条锦鸡儿中克隆了1个CDPK类基因的CDS,命名为CkCDPK。通过测序和生物信息学分析,结果显示,该基因包含1710bp的开放阅读框,编码570个氨基酸,分子质量为64.03ku,蛋白质等电点(PI)为9.12。结构分析显示,CkCDPK含有N端可变区、蛋白激酶区、4个EF手型结构和类似钙调素等结构域。利用实时荧光定量PCR检测了CkCDPK基因在不同逆境胁迫下的表达量,结果显示,NaCl胁迫和干旱胁迫下该基因的表达量呈现出单峰趋势,其中,盐胁迫6h、干旱胁迫4h表达量最高,外源ABA诱导下该基因的表达量基本呈逐渐上升趋势。表明该基因可能参与了柠条锦鸡儿的抗逆性调控。构建植物超表达载体pCAMBIA3301-CkCDPK,可为进一步研究CkCDPK基因在柠条锦鸡儿抗逆性调控中的作用奠定基础。

柠条锦鸡儿CkGR基因克隆及功能分析

通过RACE技术从柠条锦鸡儿中克隆得到一个新的GR基因,全长2 122 bp,包括5'非翻译区(5'-UTR)57 bp,3'非翻译区(3'-UTR)415 bp,开放阅读框(ORF)1 650 bp,编码550个氨基酸,推测的蛋白质分子量为59.2k Da,理论等电点为8.2,命名为Ck GR。Ck GR与鹰嘴豆Ca GR的同源性较高,为90.1%。利用染色体步移法克隆得到Ck GR起始密码子ATG上游648 bp的启动子序列,Plant CARE软件分析表明,该序列具有启动子的基本元件CAAT-box和TATA-box以及多种与逆境胁迫相关的顺式调控元件。实时荧光定量PCR分析表明,Ck GR在柠条锦鸡儿的根、茎和叶中均有表达,没有组织特异性;Ck GR的表达受低温、高盐和干旱胁迫的诱导,表明Ck GR在柠条锦鸡儿适应低温、高盐和干旱胁迫的过程中发挥作用。

柠条锦鸡儿CkP5CS基因克隆及表达特性分析

旨在克隆柠条锦鸡儿Δ~1-吡咯啉-5-羧酸合成酶基因(P5CS)的全长cDNA序列,研究其组织表达特异性,分析CkP5CS基因在低温、高盐和PEG处理下的表达情况,以阐明CkP5CS基因在柠条锦鸡儿中的生物学功能。利用RACE技术克隆P5CS基因cDNA序列,并对其全长基因进行生物信息学分析;构建系统发育树,研究其与相似序列的同源性;利用实时荧光定量PCR方法,分析CkP5CS基因的组织表达特异性以及在低温、高盐和渗透胁迫下的表达情况。结果显示,克隆得到柠条锦鸡儿P5CS基因全长c DNA序列,命名为CkP5CS,全长2 604 bp,包括5'非翻译区(5'-UTR)102 bp,3'非翻译区(3'-UTR)363 bp,开放阅读框2 139 bp,能编码712个氨基酸,预测蛋白质分子量为76.8 kD,理论等电点为8.6,二级结构主要包括α-螺旋、延伸链和无规则卷曲。多序列比对和系统进化分析表明,柠条锦鸡儿P5CS与白刺花SdP5CS、大豆GmP5CS、豇豆VuP5CS同源性较高,分别为85.1%、84.2%和82.6%。实时荧光定量PCR结果显示,CkP5CS在柠条锦鸡儿的根、茎和叶中均有表达,没有组织特异性;同时,该基因的表达受低温、高盐和渗透胁迫的诱导。克隆得到柠条锦鸡儿CkP5CS基因,其在柠条锦鸡儿适应低温、高盐和渗透胁迫过程中发挥作用。