水杨酸对NaHCO_(3)胁迫下黄芪幼苗生长及生理特性的调节效应

【目的】探究外源水杨酸(SA)缓解盐碱胁迫黄芪种子萌发及幼苗生长生理特性的效应。【方法】以蒙古黄芪为供试材料,利用100 mmol/L的NaHCO_(3)溶液模拟盐碱胁迫,分别设置0.1,0.2,0.3,0.4,0.5 mmol/L外源SA对黄芪种子萌发、幼苗生长进行调节,并以未加SA无NaHCO_(3)胁迫处理为对照1(CK1),未加SA及NaHCO_(3)胁迫为对照2(CK_(2)),测定蒙古黄芪种子萌发指标、幼苗生长及生理指标。【结果】NaHCO_(3)胁迫下(CK_(2))黄芪种子萌发、幼苗生长及生理受到明显抑制,与CK1相比,黄芪种子发芽率、发芽势、发芽指数和活力指数分别降低93.96%、93.45%、97.26%、97.50%,幼苗株高、单株鲜重、根系活力分别降低35.97%、37.61%、72.65%;超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性及可溶性蛋白、脯氨酸(Pro)受到不同程度的抑制,叶片质膜透性、丙二醛(MDA)含量显著增加;与CK_(2)相比,添加不同浓度外源SA后蒙古黄芪种子萌发及幼苗生长显著提高(P<0.05),叶片质膜透性及膜脂过氧化产物MDA含量分别降低了40.21%~56.56%、20.09%~46.39%,抗氧化酶SOD、POD、CAT活性及根系活力得到不同程度增强,渗透调节物质游离脯氨酸、可溶性蛋白积累量显著提高了7.63%~86.95%、5.51%~41.75%。【结论】外源SA可以有效调节NaHCO_(3)胁迫对黄芪种子萌发及幼苗生长的抑制,缓解渗透压并降低膜脂过氧化对黄芪幼苗带来的损伤,增强黄芪种子及幼苗对盐碱胁迫的适应性及抗盐碱能力,其以0.3 mmol/L缓解效果最佳。

珍稀濒危植物沙冬青Ammopiptanthus mongolicus(Maxim.)(chengf.)组织培养再生植株的研究

本文对珍稀濒危植物沙冬青的组织培养进行了研究。采用萌发 12 d的无菌苗的子叶和胚轴接种于含不同激素成分的 MS培养基上 ,经过 1次诱导获得了再生植株。简化了培养程序 ,缩短了培养时间。

Effects of the Extract of Ammopiptanthus mongolicus cheng f. (JA1) on Induction of Apoptosis of HepG2 in vitro and Its Molecular Mechanisms

Objective To study the effects and the mechanisms of extract from a leguminous plant （Ammopiptanthus mongolicus cheng f.） （JAl ） in northwest China on inducing apoptosis and inhibiting proliferation of HepG2 hepatocarcinoma cell in vitro. Methods The HepG2 cell line was used as target cells. The effect of 3A 1 on HepG2 cell growth was detected by microculture tetrazolium assay （MTr）, flow cytometry assay, DNA agarose gel electrophoresis and transmission electronic microscopy. The expressive effect of the wt-p53 in HepG2 cells was analyzed with p53 protein test-reagent. Results JAl not only had significant anti-proliferative effects depending upon time and dosage, but also induced apoptosis of HepG2 cells. Apoptotic typical morphological changes were observed in JAl-treated HepG2 cells under transmission electronic microscope, ＂Sub-G 1＂ phase peak occurred in flow cytometry and DNA ＂ladder＂ was found in DNA agarose gel electrophoresis. The expression of the wt-p53 increased in vitro, and 3Al-treated HepG2 and the positive cell percentage of the wt-p53 protein also increased. Conclusions JAl could obviously induce apoptosis and inhibit proliferation of HepG2 cells in vitro, and these effects are closely related with the increase of wt-p53 expression. JAl can be used as a good source of medicinal plant for the treatment of hepatocarcinoma.

Effects of cold-hardening on compatible solutes and antioxidant enzyme activities related to freezing tolerance in Ammopiptanthus mongolicus seedlings

Cold acclimation is associated with many metabolic changes that lead to an increase of freezing tolerance. In order to investigate the biochemical process of cold acclimation in Ammopiptanthus mongolicus, seedlings were acclimated at 2℃ under 16-h photoperiod （150 μmol·m^-2·s^-1 photosynthetically active radiation） for 14 d. Freezing tolerance in seedlings increased after 14 d of cold-hardening. Contents of protein, proline and solute carbohydrate in cotyledon increased after cold acclimation. Patterns of isozymes of superoxide dismutase （SOD）, peroxidase, catalase and polyphenol oxidase （PPO） were investigated. The activities of SOD, peroxidase and PPO in cold acclimated plants were increased during cold-hardening. We deduced that compatible solutes and antioxidant enzymes play important roles in development of freezing tolerance during cold acclimation in this evergreen woody plant.

Functional analysis of cold-inducible cDNA clones in the legume Ammopiptanthus mongolicus

Ammopiptanthus mongolicus is the only evergreen broadleaved shrub endemic species to the Alashan Desert, northwest sand area of China, and can survive at -30℃ or even lower temperature in winter. A modified solid-phase subtraction hybridization technique was developed to isolate and screen cDNAs, whose transcripts increased in cold-treated A. mongolicus seedlings. Sequence analysis of the screened clones indicated that 11 clones had coding regions, with four of them containing complete open reading frame. And nine of the 11 clones shared various degrees of homology with the genes found in the GenBank database and the other two were unidentified sequences. Sequence data further revealed that these accumulated transcripts encode three low molecular weight proteins, a late-embryogenesis protein and two cold acclimation responsive proteins; two photosynthesis-related proteins, photosystem I subunit II precursor (PsaD) and photosystem II oxygen-evolving complex 33kDa subunit (OEC33); a protease inhibitor; an adenosine triphosphatase and a 14-3-3 related protein. Analysis on the function of these proteins indicated that the low molecular weight proteins were associated with water holding ability of cytoplasm; photosynthesis-related proteins participated in the adjustments of photosynthetic apparatus to resist photoinhibition; 14-3-3 related protein could interact with adenosine triphosphatase to enhance ATPase activity and energy metabolism; and protease inhibitor involved in the prevention of unwanted cell death was caused by reactive oxygen species. We suggest that cold acclimation with low light intensity in A. mongolicus is a more complex interaction of low temperature, light, energy and signal than that assumed previously.

Susceptibility Test of Culter mongolicus to Several Kinds of Common Drugs

[ Objective ] This study aimed to provide references for reasonable drug administration in Culter mongolicus and promote the development of its breeding industry. [Method] C. mongolicus was administrated with cypermethrin, mebendazole, trichlorfon, cupric sulfate, trichloroisocyanuric acid, povidone iodine and phoxim, respectively, to analyze its susceptibility to different drugs. [ Result] C. mongolicus was very sensitive to cypermethrin; the median tolerance limit （TLM） at 3,8 h was 0. 072mg/L, minimum lethal concentration was 0. 063 - 0. 069 mg,/L, safe concentration was 0. 021 6 mg/L. C. mongolicus was insensitive to mebendazole, trichlorfon, cupric sulfate, trichloroisocyanuric acid and povidone iodine. According to safe concentration of various drugs, C. mongolicus exhibited higher susceptibility to cypermethrin than to phoxim. [ Conclusion] This study laid a solid foundation for better protection and development of C. mongolicus resources.

丛枝菌根真菌(AMF)对蒙古沙冬青幼苗的促生特性及作用机制

为解决蒙古沙冬青(Ammopiptanthus mongolicus)幼苗根系不发达、再生性差、移栽造林成活率低的技术难题及其资源的有效扩繁和保存,试验采用丛枝菌根真菌(AMF)不同接菌方式处理蒙古沙冬青实生幼苗根系,分析AMF对其侵染状况及植株生长和生物量的变化,根系及叶片生理特性、根际土壤酶活性变化及其与幼苗生长和生物量变化的关联性。研究旨在探讨根系菌根化作用对蒙古沙冬青幼苗的促生长及作用机制。结果表明:丛枝菌根真菌(根内根孢囊霉Rhiaophagus intraradice,摩西斗管囊霉Funneliformis mosseae)可有效侵染蒙古沙冬青幼苗根系并与其根系构建互惠共生体,不同菌种与宿主植物的亲和程度存在差异。(1)单接菌和混合接菌与未接菌(CK)相比生长量均有所提高,主要表现在植物株高(34.7%~47.3%)和根长(32.7%~72.9%)相对较大。其中混合接菌较单接菌处理显著增加了总根投影面积、总根体积、根冠比、根干重及根表面积、植株总生物量和苗木质量指数。(2)接种AMF,特别是接种R.i+F.m幼苗根系活力、根系阳离子交换量及根系可溶性蛋白含量较对照显著提升338.7%、177.2%和240.4%,根系硝酸还原酶及碱性磷酸酶活性也显著高于单一接菌(R.i、F.m)及未接菌处理。同时该处理下幼苗叶净光合速率和可溶性蛋白含量较对照显著提高237.5%和54.3%。(3)3种接菌方式均可显著提升蒙古沙冬青幼苗根际土壤脲酶、碱性磷酸酶和蔗糖酶活性,且3种土壤酶均在混合接菌时活性提升速率最为显著,分别较对照激增564.7%、145.8%和154.3%。综合以上指标分析,AMF通过侵染蒙古沙冬青根系刺激幼苗根际土壤酶的产生和分泌并促进其根系阳离子交换及活力提升,有助于增强根系代谢过程中酶促作用及蛋白等营养物质的积累,同时影响和改善了幼苗光合代谢和营养储存,从而促进沙冬青根系及幼苗生长和生物量累积。

Comparative study of the nutritional composition and toxic elements of farmed and wild Chanodichthys mongolicus

Information of the difference in quality between farmed and wild fish is central to better ensuring fish products produced in aquaculture meet regulatory and consumer requirements.Proximate composition,amino acid and fatty acid profiles,and toxic elements contents of farmed and wild Chanodichthys mongolicus were established and compared.Significantly higher crude protein content while lower moisture content in farmed fish compared to wild fish were observed(P<0.05).The percentages of total amino acids(TAA),total essential amino acids(TEAA),total non-essential amino acids(TNEAA)and total delicious amino acids(TDAA) in farmed fish were all significantly higher than those in the wild equivalent(PO.05).Compared to the FAO/WHO recommended reference values,the ratios of TEAA/TAA(39.84%-40.33%) were comparable to 40%and TEAA/TNEAA(66.22%-67.60%) were above 60%.Fatty acid profiles in both fanned and wild C.mongolicus were dominated by monounsaturated fatty acid(MUFA),with farmed fish contained much more MUFA content compared to wild counterpart(P<0.05).Notably,wild fish exhibited significantly higher levels of total polyunsaturated fatty acid(PUFA) including eicosapentaenoic acid(EPA) and docosahexaenoic acid(DHA) than farmed fish(P<0.05).The EPA(C_(20:5n3))and linoleic acid(C_(18:2n6)) were the predominant PUFA in wild and fanned C.mongolicus,respectively.Moreover,farmed fish displayed an overall lower toxic element levels(As,Cd,Pb and Hg) in comparison with wild fish,and both were far lower than the established limit standard.In conclusion,our results suggest that the nutritional quality of farmed C.mongolicus was inferior to their wild counterpart with respect to fatty acids nutrition,and therefore further studies should focus on the improving C.mongolicus diet in order to enhance the overall nutritional composition.

Determining oxygen consumption rate and asphyxiation point in Chanodichthys mongolicus using an improved respirometer chamber

Knowledge of oxygen consumption rates and asphyxiation points in fish is important to determine appropriate stocking and water quality management in aquaculture.The oxygen consumption rate and asphyxiation point in Chanodichthys mongolicus were detected under laboratory conditions using an improved respirometer chamber.The results revealed that more accurate estimates can be obtained by adjusting the volume of the respirometer chamber,which may avoid system errors caused by either repeatedly adjusting fish density or selecting different equipment specifications.The oxygen consumption rate and asphyxiation point of C.mongolicus increased with increasing water temperature and decreasing fish size.Changes in the C.mongolicus oxygen consumption rate were divided into three stages at water temperatures of 11-33℃:(1) a low temperature oxygen consumption rate stage when water temperature was 11-19℃,(2) the optimum temperature oxygen consumption rate stage when water temperature was19-23℃,and(3) a high temperature oxygen consumption rate stage when water temperature was >27℃.The temperature quotients(Q_(10)) obtained suggested that C.mongolicus preferred a temperature range of19-23℃.At 19℃,C.mongolicus exhibited higher oxygen consumption rates during the day when the maximum values were observed at 10:00 and 14:00 than at night when the minimum occurred at 02:00.

Analysis on Volatile Components of Flowers and Leaves of Thymus mongolicus by SPME-GC/MS

[Objectives] To analyze volatile components of flowers and leaves of Thymus mongolicus. [Methods]Volatile components of collected T. mongolicus flower and leaves were separated by headspace solid-phase microextraction( SPME) and identified by gas chromatography/mass spectrometry( GC/MS) and normallized by peak area. [Results] A total of 24 and 14 compounds were identified from flowers and leaves of T. mongolicus in the total ion chromatogram,accounting for 99. 573% and 97. 187% of the total peak area,respectively. Main components of flowers and leaves of T. mongolicus include phenols and terpenes,and thymol accounts for 35. 38% and 49. 13% of flowers and leaves of T. mongolicus,respectively. [Conclusions] SPME-GC/MS can be applied in analyzing volatile components of flowers and leaves of T. mongolicus,to provide basis for further development and utilization of T. mongolicus.

Cold acclimation induced accumulation of phenolic compounds and freezing tolerance in Ammopiptanthus mongolicus

Ammopiptanthus mongolicus, the only freezing tolerant evergreen broad-leaved shrub, local species of the Alashan desert, northwest sand area of China, can survive -30°C or even lower temperature in winter. In the present study, the secondary products phenolics in A. mongolicus cotyledons were determined to study the effects of phenolics on cold tolerance. Cytochemical localization of phenolics in cotyledon cells was observed by electron microscopy and the content of phenolic compounds was assayed by spec-trophotometric measurement. The results showed that the freezing tolerance of A. mongolicus seedlings increased after acclimation at 2-6°C for 14 days, which accompanied the increase of the content of phenolic compounds in cotyledons. Cytochemical observation showed that phenolic deposits were mainly localized in vacuoles and in close proximity to tonoplast, and also in the cytoplasm. The amount and the size of phenolics droplets increased obviously in cytoplasm and vacuoles after cold acclimation, predominantly ag-gregated along membranes of vacuoles and tonoplast. No phenolic deposits were found in cell walls. As hydrogen-or elec-tron-donating agents, phenolics may protect plant cells against reactive oxygen species formed during chilling or freezing stress and improve the freezing tolerance of cold-acclimated A. mongolicus seedlings.

沙冬青（Ammopiptanthus mongolicus）小孢子发育的超微结构研究

本文对沙冬青小孢子发育的超微结构进行了观察，主要结果为：１）小孢子母细胞早期壁上有胞间连丝，随着发育，壁上出现了胞质通道．母细胞中内质网、线粒体和质体丰富，细胞处于明显的代谢活跃状态．四分体时期内质网消失．２）在小孢子发育各期质体和线粒体一直存在，细胞质中分布着小液泡，质膜上有泡状内陷．小孢子核靠壁后发生有丝分裂形成两个大小悬殊的细胞，此时，可见到大量的质体集中分布在营养细胞质中靠近生殖细胞壁的附近．３）在小孢子早期，绒毡层中内质网、线粒体和质体丰富，内质网持续产生大量小泡并在其中沉积电子致密物形成球状体（Ｏｒｂｉｃｕｌｅ）．小孢子后期，绒毡层细胞中内质网消失，出现大量的多泡体、球状体和脂体，绒毡层开始解体．在２－细胞花粉时绒毡层的解体产物大量流入花药腔中．

Molecular cloning and functional analyses of low-temperature induced genes from Ammopiptanthus mongolicus

Studies on the cold-responsive genes and cold signaling of woody species drop far behind in comparison to herbaceous plants.Due to similar lignified structure,perennial characteristic,and enhanced tolerance,it seems much easier to find strongly antifreeze genes and obtain effective results in transgenic woody plants.In this study,Ammopiptanthus mongolicus,an evergreen,broadleaf and cold-resist leguminous shrub growing in the desert of Inner Mongolia,was used as a material for low-temperature induced gene isolation.Through differential expression analysis induced by low-temperature,thirteen up-regulated cDNAs were identified.One of them,AmEBP1,(accession number:DQ519359)confers enhanced cold-tolerance to both transgenic E.coli and transgenic Arabidopsis.Results suggest that AmEBP1 can stimulate the synthesis of ribosome and the dephosphyration of the α-subunit of initiation factor 2(eIF2α),and subsequently promote the translation process.By which the transgenic plants obtained increased cold-resistant ability.

百里香内生真菌塞缪尔斯木霉Trichoderma samuelsii 2-63菌株的鉴定及其抗真菌活性

植物病原真菌是导致作物产量损失和品质下降的重要因素之一,因此,开发低毒高效的抑菌剂一直是研究的热点。本研究从百里香植株茎中分离得到1株内生真菌2-63菌株,通过形态学结合分子生物学鉴定其为塞缪尔斯木霉Trichoderma samuelsii。平板拮抗活性表明:T.samuelsii 2-63菌株对8种植物病原真菌的拮抗活性均达70.0%以上,其中对枸杞黑霉病菌Alternaia alternata拮抗活性最高可达87.1%;平板共培养试验表明:T.samuelsii 2-63菌株挥发性有机化合物(VOCs)也表现出对病原菌不同程度的抑制活性,抑制率在31.0%~68.5%;但T.samuelsii 2-63菌株液体发酵液和粗提物的抑菌活性较低。抑菌活性结果结合气相色谱-质谱联用(GC-MS)分析试验结果表明:T.samuelsii 2-63菌株抑菌活性的关键成分为VOCs,其主要组分为6-戊基-2H-吡喃-2-酮(6-PP)。探索性试验表明:固体发酵条件下,大米培养基比大麦培养基更利于6-PP的产生,最大产量为38.6 mg/mL;浅层液体培养条件下,6-PP随着培养时间的增加出现先上升后下降的趋势,在培养第15天时产量最大,为0.1 mg/mL。该研究结果为基于木霉菌的微生物菌剂和农药的开发提供了一定的理论依据和试验基础。

天然沙冬青器官生态化学计量特征对异质生境的响应

以我国重点保护的珍稀濒危植物且是西北荒漠地区唯一常绿阔叶灌木沙冬青(Ammopiptanthus mongolicus)为研究对象,分析异质性荒漠生境(固定沙地、半固定沙地、石质沙地、洪积砾石坡地和盐碱滩地)对沙冬青叶、茎、根、花和种子生态化学计量学特征的影响,了解其生长的适应机制和生存策略。结果表明:沙冬青各器官碳(C)含量表现为叶>茎>根>种子>花,而氮(N)、磷(P)、钾(K)营养元素含量均表现为种子>花>叶>根>茎,N、P、K三大营养元素在繁殖器官中的富集,可以促进沙冬青由营养生长向生殖生长的转变,反映了资源分配模式和植物生长策略。沙冬青各器官的碳氮磷钾含量和各元素化学计量比在不同生境条件下变化较复杂,其中在沙质土壤(固定和半固定沙地)具有更高的C、N、P储存能力,而K在砾质土壤(石质沙地和洪积砾石坡地)含量较高,且砾质荒漠沙冬青的C∶N、C∶P及N∶P均显著高于沙质土壤,表明沙冬青在石质沙地和洪积砾石坡地有更高的养分利用效率。各器官元素含量相关分析表明器官间的养分协同性远高于器官内部。化学计量比C∶N、C∶P、C∶K均表现为:茎>根>叶>花>种子,N∶P为根>叶>茎>花>种子,N∶K为茎>根>叶>种子>花,K∶P为叶>根>种子>茎>花,各器官中的C∶N和N∶P比在各生境间相对较稳定,而K∶P比变化巨大,N∶P与P含量呈显著的负相关(P<0.05),与N含量的相关性不显著(P>0.05),说明P作为敏感性元素决定了沙冬青体内N∶P比值的变化,且不同生境叶片N∶P比值均大于16,说明沙冬青生长主要受P限制。器官与生境对沙冬青化学计量特征都有影响,生境对C含量和N∶K、K∶P的影响较大,器官对N、P、K含量及C∶N、C∶P、C∶K、N∶P的影响较大。除P元素外,各元素含量及其化学计量比均受器官和生境交互作用影响。了解植物根-茎-叶-花-种子的整体资源权衡和协变策略,为沙冬青物种资源的保护和开发提供理论依据。

长白山区天然蒙古栎林立地类型划分

基于2014年吉林省第八次森林资源连续清查数据,调查490块天然蒙古栎林样地。采用主成分分析和聚类分析方法对长白山区天然蒙古栎林立地类型进行划分。主成分分析结果表明:立地因子对立地类型划分的影响顺序为坡向>坡位>坡度>土层厚度>海拔>腐殖质厚度。当聚类系数为0.6683时,可将490块天然蒙古栎林样地划分为4类:Ⅰ类立地因子多为阴坡、中坡、陡坡、薄土;Ⅱ类立地因子多为半阴坡、上坡、斜坡、薄土;Ⅲ类立地因子多为阳坡或半阳坡、上坡、斜坡、中土;Ⅳ类立地因子多为阳坡、上坡、平缓坡、中土,最适合天然蒙古栎林生长的是Ⅲ类立地类型。

沙冬青CBF/DREB1转录因子cDNA的克隆及序列分析

CBF/DREB1(C-repeat binding factor/dehydration resistance element binding protein 1)是一类抗逆相关的转录因子,它们的表达可以激活下游一系列抗逆应答基因的表达,从而提高植株的抗逆性。沙冬青(Ammopiptanthus mongolicus)主要生长在中国西北荒漠和半荒漠地带,是典型的旱生资源植物,具有突出的抗寒、抗旱、耐盐碱特性。本文以沙冬青为实验材料,利用RT-PCR和RACE技术克隆了沙冬青CBF/DREB1基因cDNA序列,并对其进行了序列分析。结果表明:沙冬青CBF/DREB1转录因子基因序列全长为991bp,包括624bp的开放阅读框(ORF),起始密码子ATG,终止密码子TAA,114bp的5'UTR和253bp的3'UTR,以及加尾信号AATAAA及poly(A)11。生物信息学预测其演绎的氨基酸序列具有AP2结构域,且AP2结构域的两端拥有CBF家族特有的PKK/RPAGRxKFxETRHP和DSAWR两段短多肽序列,属于CBF家族。沙冬青CBF/DREB1转录因子的克隆为进一步研究植物抗逆和获得抗性基因提供了新的候选基因,也为进一步从分子水平上验证其功能,深入理解植物适应干旱、低温和高盐机制奠定基础。

蒙古沙冬青根围AM和DSE真菌与土壤因子的相关性研究

于2012年6月对银川、沙坡头、民勤3个样地的蒙古沙冬青（Ammopiptanthus mongolicus）根围采集5个土层（0~10、10~20、20~30、30~40、40~50cm）的土样和根样,研究了蒙古沙冬青根际AM和DSE真菌空间分布以及土壤因子的生态作用,以阐明沙冬青与土壤真菌的共生关系,为利用共生真菌资源促进沙冬青生长和荒漠植被恢复提供依据。结果表明：（1）沙冬青根系能被AM和DSE真菌高度侵染,共生程度和生态分布具有明显空间异质性。（2）沙坡头样地AM真菌总定殖率最高,民勤DSE定殖率最高,但不同样地菌丝、泡囊、丛枝和定殖强度无显著差异;同一样地不同土层AM真菌总定殖率和DSE定殖率无显著差异。（3）相关性分析表明,AM真菌总定殖率与土壤速效P显著正相关;泡囊定殖率与有机质显著正相关,与速效P极显著正相关;定殖强度与总球囊霉素（TEG）显著正相关,与碱解N和速效K极显著正相关;DSE定殖率与易提取球囊霉素（EEG）极显著正相关,与碱解N显著正相关。（4）通径分析表明,土壤有机质、EEG和速效K是通过直接作用影响AM真菌总定殖率和DSE定殖率。

改良Trizol法提取高质量蒙古沙冬青总RNA

以木本植物蒙古沙冬青的叶片和根为材料,采用改良Trizol法提取总RNA。结果表明,与传统Trizol法相比,改良Trizol法能有效去除提取过程中残留的异硫氰酸胍、β-巯基乙醇以及植物本身的糖、酚类物质,获得高质量总RNA。改良Trizol法提取总RNA技术已被成功应用于蒙古沙冬青转录组数据库的建立。该方法可为木本植物提取高质量RNA提供借鉴。

蒙古沙冬青冷冻胁迫SMART cDNA文库的构建及序列分析

以强抗逆植物蒙古沙冬青(Ammopiptanthus mongolicus)为材料,用SMART技术构建了其在冷冻胁迫下的全长cD-NA文库。原始文库滴度为9.44×106pfu/ml,重组率为98.3%,插入片段长度在0.5～2.5kb,平均达到1kb。扩增文库滴度为1.98×1010pfu/ml。从原始文库中随机挑取480个阳性克隆进行序列分析,获得348个Unigene,其中有些是逆境应答相关基因和新基因。此外,以扩增文库为模板,经PCR方法克隆到读码框为1083bp的沙冬青类RD22基因。结果表明所建文库质量较高,可以满足后续研究的要求。