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1.
为了分离与鉴定巨龙竹速生巨大相关基因,以巨龙竹当年所发新叶为材料,用植物叶RNA抽提试剂盒提取了总RNA;采用C reatorTMSM ARTIMcDNA L ibrary construction k it所提供的方法,合成及纯化cDNA,并将cDNA连接到质粒载体上,通过C aC l2法将重组质粒转化到DH 5a中,成功构建了巨龙竹cDNA文库。文库容量为1.04×105,PCR检测表明大多数插入片段均大于1 000 bp,表明构建的巨龙竹cDNA文库质量较高,为进一步进行EST测序和全长基因克隆打下了基础。 相似文献
2.
胡杨根、茎、叶、愈伤组织总RNA提取的研究 总被引：1，自引：0，他引：1
胡杨根、茎、叶、愈伤组织等器官和组织的总RNA高质量提取是进行以胡杨RNA为基础的分子生物学研究的基础。以2 a生胡杨实生苗根系为材料,用CTAB-PVP提取缓冲液裂解细胞并变性蛋白,经氯仿/异戊醇(24∶1,v/v)抽提后,用3种不同的沉淀方法获得胡杨根系总RNA,电泳和定量测试等分析提取质量。结果表明,LiCl沉淀法结合70%乙醇洗涤沉淀两次获得的的胡杨根系总RNA,条带清晰、亮度高,且没有明显的DNA污染;actin基因片段设计引物对胡杨根系总RNA进行RT-PCR扩增,扩增片段与预期大小一致,表明该RNA可用于逆转录合成cDNA,提取物RNA中的Li 残留对逆转录酶活性的抑制作用不明显;该方法用以提取2 a生胡杨实生苗茎、叶及愈伤组织总RNA,均获得高质量总RNA。CTAB-PVP-LiCl(附70%乙醇洗涤RNA沉淀2次)的方法可用于胡杨根、茎、叶和愈伤组织总RNA的提取。 相似文献
3.
目的:探讨驱虫斑鸠菊对人 A375黑素瘤细胞株分泌酪氨酸酶活性、黑素合成以及细胞内酪氨酸酶基因表达水平的影响。方法:采用酶学方法研究驱虫斑鸠菊对酪氨酸酶活性的影响;采用475 nm 比色法测定黑素含量;采用 TRIzol 试剂盒提取总 RNA,再用半定量逆转录聚合酶链反应扩增 cDNA,检测酪氨酸酶基因表达水平。结果:驱虫斑鸠菊可以增强 A375人黑素瘤细胞酪氨酸酶活性和黑色素合成能力,还可增强酪氨酸酶基因表达。结论:驱虫斑鸠菊可能从基因水平增强酪氨酸酶活性,促进黑素合成。 相似文献
4.
樟叶越桔叶芽总RNA提取方法比较研究 总被引：2，自引：0，他引：2
本项实验采用改良CTAB、改良SDS、RNApure Plant Kit和Plant RNA Kit 4种方法提取樟叶越桔叶芽总RNA,以期获得适合于樟叶越桔叶芽高质量总RNA的最佳提取方法。总RNA纯度和完整性分别用紫外分光光度计法和琼脂糖凝胶电泳法检测。结果表明,采用改良CTAB和Plant RNA Kit法获得的RNA完整性好,纯度高,A260/A280值分别为2.07和2.08,28S和18S rRNA条带清晰,且前者的亮度约为后者的2倍,无弥散现象。将改良CTAB和Plant RNA Kit法获得的总RNA进行RT-PCR扩增,成功克隆获得了樟叶越桔花色苷5-芳香族酰基转移酶基因长316 bp的cDNA序列。证明改良CTAB和Plant RNA Kit法是樟叶越桔叶芽高质量总RNA的最适提取方法。 相似文献
5.
为了筛选最适的菠萝蜜总RNA提取方法,采用Trizol提取法、Tris-硼酸提取法和试剂盒提取法提取菠萝蜜叶片和种子总RNA,通过紫外分光光度计检测、琼脂糖凝胶电泳、RT-PCR检测以及Agilent2100检测,对提取结果进行比较分析。结果表明:采用这3种方法均能从菠萝蜜叶片和种子中提取到总RNA,Tris-硼酸提取法较其他2种方法所提取的总RNA产率高,但杂质较多;采用Tris-硼酸提取法能提取完整性较好的总RNA,28S、18S和5S清晰可见,但提取质量较试剂盒提取法差;试剂盒提取法省时省力,有明显的28S和18S条带,A260/A280分布在1.92~1.98,A260/A230分布在2.04~2.09,叶片总RNA产率42.3ng/μL,种子总RNA产率15.6ng/μL。RT-PCR检测结果表明在约200bp处存在清晰整洁的电泳条带,Agilent2100检测结果显示杂峰极少,28S峰积分面积约是18S峰积分面积的2倍。经综合评定认为,试剂盒提取法是适合菠萝蜜叶片和种子总RNA提取的方法。 相似文献
6.
7.
《中南林业科技大学学报(自然科学版)》2016,(7)
试验以天女木兰叶芽、幼叶、成熟叶、花芽、花瓣、种子为材料,分别采用Trizol法、改良Trizol法、CTAB-异丙醇法、CTAB-Na Ac和无水乙醇法、CTAB-Li Cl法、天根试剂盒法对其各器官进行总RNA的提取。总RNA的完整性、纯度和浓度等提取效果分别通过琼脂糖凝胶电泳和酶标仪进行检测。结果表明:叶芽的总RNA最适合用改良Trizol法进行提取,花瓣和花芽的总RNA最适合用改良的CTAB法提取,幼叶的总RNA可选择试剂盒法,Trizol可用于成龄叶片的提取,而天根试剂盒最适合天女木兰种子总RNA的提取。通过不同提取方法获得的高质量总RNA经RT-PCR检测表明:可以直接用于后续分子克隆和基因表达分析等分子生物学实验。 相似文献
8.
枣RNA提取方法的比较 总被引：3，自引：1，他引：2
为给在分子水平上对枣进行研究提供参考,通过采用分光光度法和琼脂糖凝胶电泳法检测提取样品,比较CTAB-异丙醇法、CTAB-LiCl法、Trizol-异丙醇法和Trizol-LiCl法等4种RNA提取方法对枣RNA提取的影响。结果表明,CTAB-异丙醇法提取的枣RNA纯度和完整度均较高,杂质较少,DNA残留少,降解不明显,可以用在枣枝条韧皮部、果实、嫩叶和老叶等器官和组织的RNA提取上,应用该方法所提取的RNA可以作为cDNA合成和基因克隆的模板。 相似文献
9.
山核桃花芽总RNA提取方法研究 总被引：4，自引：0，他引：4
以山核桃的花芽为材料,利用改良CTAB法、异硫氰酸胍法和改良热酚法提取其总RNA,并对RNA产率、纯度、电泳图谱和RT-PCR进行分析,结果表明:改良CTAB法明显优于其它两种提取方法,用它提取的28S rRNA、18S rRNA条带清晰、稳定,无降解,OD260/OD280值保持在1.9～2.0,产率相对较高;经RT-PCR检测发现,改良CTAB提取的RNA能被成花基因特异引物扩增出清晰的条带,说明该法完全适合于山核桃花芽总RNA的提取,并可用于Northern杂交、cDNA文库构建等分子生物学操作. 相似文献
10.
《浙江林业科技》2016,(5)
采用CTAB法、CTAB-Li Cl法、CTAB-Trizol法、Trizol法、天根RNA提取试剂盒及百泰克RNA提取试剂盒提取草珊瑚(Sarcandra glabra)叶片总RNA,并通过微量紫外分光光度计、电泳检测等对提取的结果进行了分析比较。结果表明:CTAB-LiCl法和天根RNA提取试剂盒比较适合提取草珊瑚叶片的总RNA。这两种方法提取的总RNA的完整性和纯度较高,无明显的蛋白质及其他杂质污染,OD_(260)/OD_(280)的值在1.9~2.1,OD260/OD230在2.0~2.4。获得质量高、完整性好、纯度高的总RNA,为后续构建cDNA文库以及相关基因的克隆提供了试验基础。 相似文献
11.
从黑龙江省双城林场采集健康的分月扇舟蛾2龄幼虫,将室内保存一年的分月扇舟蛾虫尸研碎,分离提纯得到纯净的颗粒体病毒。将病毒悬液稀释为1..58×103PIB·mL-1,1.58×104PIB·mL-1,1.58×105PIB·mL-1,1.58×106PIB·mL-1,1.58×107PIB·mL-1,1.58×108PIB·mL-1,1.58×109PIB·mL-17种浓度,采集新鲜的杨树叶分别浸入上述不同浓度的悬液,对采集的健康幼虫进行接毒。9天实验结果表明,浓度为1..58×103PIB·mL-1时校正死亡率最小(7.32%),浓度是1.58×109PIB·mL-1时校正死亡率最大(97.36%),浓度对数和死亡几率值的回归直线方程为y=1.946 0.558x,LC50为2.97×105PIB·mL-1。1.58×105PIB·mL-1,1.58×106PIB·mL-1,1.58×107PIB·mL-1,1.58×108PIB·mL-1,1.58×109PIB·mL-1w所对应的半致死时间(LT50)分别为8.55d,6.89d,5.9d,4.65d,4.08d。随着浓度的增大,LT50逐渐缩短,说明该病毒的毒力较强,作为杀虫剂具有很好的应用潜力。 相似文献
12.
An integrated model for predicting maximum net photosynthetic rate of cocksfoot (Dactylis glomerata) leaves in silvopastoral systems 总被引：1，自引：0，他引：1
Net light-saturated photosynthetic rate (Amax) of field grown cocksfoot (Dactylis glomerata L.) leaves in a radiata pine (Pinus radiata D. Don) silvopastoral system (Canterbury, New Zealand) was measured at different times under severe shade (85–95 μmol m–2 s–1 photosynthetic photon flux density, PPFD) and in full sunlight (1900 μmol m–2 s–1 PPFD). The aim was to integrate individual functions for Amax against air temperature (2 to 37 oC), water status, expressed as pre-dawn leaf water potential (ψlp) (-0.01 to −1.6 MPa), herbage nitrogen (N) (1.5 to 5.9%), regrowth duration (20 to 60 days) and time under shade (1 to 180
min) into a multiplicative model. The highest Amax value obtained was 27.4 μmol CO2 m–2 s–1 in non-limiting conditions with full sunlight. This value was defined as standardised dimensionless Amaxs = 1 for comparison of factor effects. The canopy temperature of the cocksfoot sward was up to 7.4 oC cooler than air temperature
for plants under shade. Therefore, canopy temperature was used to predict Amax. The only interaction was between time under severe shade (5% of the open PPFD) and water stress (ψlp = −0.4 to −1.3 MPa) and this was included in the model. Validation of this model indicated 78% of the variation in Amax could be accounted for using these five factors by the addition of the interaction function. This model could be used to
assist the prediction of pasture growth in silvopastoral systems through incorporation into a canopy photosynthesis model.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
13.
Zhende Yang Boguang Zhao Lin Zhu Jie Fang Luqing Xia 《Frontiers of Forestry in China》2006,1(2):190-195
Alkaloids from Sophora alopecuroids were bioassayed with Clostera anastomosis for their antifeedant and growth inhibitory effects. The antifeedant rate in choice test reached 62%–86% at the dose 2.5
mg/mL, while in non-choice bioassay the rate was only 20%–29%. In choice bioassay, the antifeedant rate increased with larval
instars of C. anastomosis and did not in non-choice experiment. The alkaloids also imposed a strong influence on the growth of C. anastomosis larvae, i.e., after feeding on the leaves treated with alkaloid, the larvae lost their weight, weight gain, and relative
growth rate (RGR) significantly when compared with the controls. In the second day after treatment with the dose at 10 mg/mL
of the alkaloid, the RGR reduced by 39.8%, and the food intake and the feces weight were respectively 57.7% and 57.4% of the
controls. The approximate digestibility (AD) increased significantly, and the efficiency in converting digested food (ECD),
and the efficiency in converting ingested food (ECI) decreased greatly after feeding the treated leaves. Moreover, the eggs
laid per female were also inhibited by this alkaloid. The significance and prospect of the alkaloids in controlling forest
insect pests were also discussed.
__________
Translated from Scientia Silvae Sinicae, 2005, 41(4) [译自: 林业科学, 2005, 41(4)] 相似文献
14.
The impacts of elevated atmospheric CO2 concentrations (500 μmol·mol−1 and 700 μmol·mol−1) on total soil respiration and the contribution of root respiration ofPinus koraiensis seedlings were investigated from May to October in 2003 at the Research Station of Changbai Mountain Forest Ecosystems, Chinese
Academy of Sciences, Jilin Province, China. After four growing seasons in top-open chambers exposed to elevated CO2, the total soil respiration and roots respiration ofPinus koraiensis seedlings were measured by a Li-6400-09 soil CO2 flux chamber. Three PVC cylinders in each chamber were inserted about 30 cm into the soil instantaneously to terminate the
supply of current photosynthates from the tree canopy to roots for separating the root respiration from total soil respiration.
Soil respirations both inside and outside of the cylinders were measured on June 16, August 20 and October 8, respectively.
The results indicated that: there was a marked diurnal change in air temperature and soil temperature at depth of 5 cm on
June 16, the maximum of soil temperature at depth of 5 cm lagged behind that of air temperature, no differences in temperature
between treatments were found (P>0.05). The total soil respiration and soil respiration with roots severed showed strong diurnal and seasonal patterns. There
was marked difference in total soil respiration and soil respiration with roots severed between treatments (P<0.01); Mean total soil respiration and contribution of root under different treatments were 3.26, 4.78 and 1.47 μmol·m−2·s−1, 11.5%, 43.1% and 27.9% on June 16, August 20 and October 8, respectively.
Foundation item: This study was supported by the Knowledge Innovation Project of the Chinese Academy of Sciences (KZCX1-SW-01) and the National
Natural Science Foundation of China (30070158).
Biography: LIU Ying (1976-), female, Ph. D. Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016, P. R. China.
Responsible editor: Song Funan 相似文献
15.
Fude Liu Ming Zhang Wenjin Wang Shuning Chen Jianwei Zheng Wenjie Yang Fengqin Hu Shuqing An 《Frontiers of Forestry in China》2009,4(1):75-84
In order to make clear the relationships between photosynthesis and leaf N, leaf P and SLA of tropical trees, and test the
differences in the relationships among life-form groups (trees, shrub-like trees and shrubs), seedlings and saplings of 101
species from a tropical montane rain forest, located in the Diaoluo Mountain of Hainan Island, were selected. The net photosynthesis
based on area and mass (A
area and A
mass), leaf nitrogen content based on area and mass (N
area and N
mass), leaf phosphorus content based on area and mass (P
area and P
mass) and specific leaf area (SLA) were measured and/or calculated. The results showed that A
area and A
mass tended to follow the order of shrubs > trees > shrub-like trees. One-way ANOVA showed that the difference in A
area between shrubs and shrub-like trees was significant (p<0.05), and for A
mass there were significant differences between shrubs and shrub-like trees and between shrubs and tree species (p<0.05). The relationships between A
area and N
mass were highly significant in all three life-form groups and for all species (p<0.0001). The correlation between A
area and P
mass was highly significant in shrubs (p = 0.0038), shrub-like trees (p < 0.0002) and for all species (p<0.0001), but not significant in trees (p>0.05). The relationship between A
area and SLAwas highly significant in shrubs (p = 0.0006), trees (p<0.0001) and for all species (p<0.0001), however this relation was not significant in shrub-like trees (p>0.05). The relationships between A
mass and leaf N and SLA were highly significant in all three life-form groups and for all species (p<0.0001). For A
mass and leaf P, there were significant correlations in tree groups (p = 0.0377) and highly significant correlations in shrub groups (p = 0.0004), shrub-like tree groups (p = 0.0018) and for all species (p < 0.0001). Stepwise regression showed that predicted A
mass values were closer to the observed values than those for predicted A
area values. Thus, it can be concluded that the relationships obtained from seedling and sapling measurements are close to those
from mature individuals; correlations between photosynthesis and N
mass, P
mass and SLA traits are significant and the relationships are stronger and more stable for A
mass than for A
area.
__________
Translated from Acta Ecologica Sinica, 2007, 27(11): 4651–4661 [译自:生态学报] 相似文献
16.
[目的]分离并量化土壤自养呼吸和异养呼吸,探讨各自贡献率及其随季节变化的动态特征。[方法]采用壕沟法和气体红外分析法,研究黄河小浪底库区山地栓皮栎人工林土壤总呼吸、自养呼吸和异养呼吸速率的季节动态变化、贡献率和环境影响因子。[结果]表明:栓皮栎人工林总土壤呼吸、自养呼吸和异养呼吸均呈夏季速率高、冬季速率低。栓皮栎土壤总呼吸、自养呼吸及异养呼吸速率与5 cm土壤温度均呈极显著指数相关,温度敏感性系数Q_(10)值大小为自养呼吸(3.40)异养呼吸(2.90)土壤总呼吸(2.45);栓皮栎土壤总呼吸、自养呼吸、异养呼吸速率与0 10 cm土壤体积含水量均显著线性相关;土壤总呼吸、自养呼吸速率与0 10 cm土壤电导率显著相关。土壤总呼吸和异养呼吸的温度敏感系数Q_(10)值均在冬季最大,夏秋季最小;而自养呼吸的Q_(10)值则呈相反的变化趋势。栓皮栎人工林自养呼吸和异养呼吸对土壤总呼吸的月贡献率为13.23%37.33%和62.67%86.76%,且自养呼吸的贡献率与土壤温度的季节变化规律相似。土壤总呼吸、异养呼吸与自养呼吸的CO2年通量分别为1 616.41、1 199.39、417.02 g·m~(-2)·a~(-1)。[结论]经过区分与定量化土壤总呼吸及其组分,确定异养呼吸为本研究区栓皮栎人工林土壤总呼吸的主要组分,作用于异养呼吸的生物与非生物因子均能显著影响整个森林生态系统表层CO_2总排放通量的大小,进一步为该研究区森林生态系统碳循环与能量流动的进一步量化研究提供参考。 相似文献
17.
[目的]为了构建松材线虫(Bursaphelenchus xylophilus)果胶酶Bxpel2基因干扰载体.[方法]通过Trizol 法提取松材线虫总RNA,反转录合成cDNA,设计带T7启动子的果胶酶Bxpel2基因引物,以cDNA为模板扩增出果胶酶Bxpel2基因片段,连接到RNA干扰载体,再以干扰载体为模板,PCR扩增出目的片段后进行测序鉴定,合成果胶酶Bxpel2基因双链RNA(dsRNA),采用RT-PCR检测松材线虫Bxpel2基因干扰后的表达情况.[结果]表明:1)提取的松材线虫总RNA完整性好,无降解;2)成功克隆出松材线虫果胶酶Bxpel2基因片段(790 bp)并将其连接至pMD19-T载体;3)以RNA干扰载体为模板合成dsRNA,浓度分别为1.313 mg·mL-1和1.152 mg·mL-1;4)RT-PCR结果显示,松材线虫经过dsRNA干扰后,Bxpel2基因表达基本受到抑制.[结论]成功构建松材线虫果胶酶Bxpel2基因干扰载体,为进一步研究Bxpel2基因在松材线虫致病过程中的作用和功能奠定了良好的基础. 相似文献
18.
Rui Katahira Hiroshi Kamitakahara Toshiyuki Takano Fumiaki Nakatsubo 《Journal of Wood Science》2006,52(3):255-260
A synthetic method for obtaining lignin oligomer that contains only the β-O-4 structure is described in detail. This method consists of three reaction steps: (1) the synthesis of t-butoxycarbonylmethyl vanillin (2), (2) the nucleophilic addition oligomerization of compound 2, and (3) the reduction of the oligomeric β-hydroxyl ester. In the first step, compound 2 was synthesized from vanillin in 96.8% yield. In the second step, compound 2 was oligomerized with commercial lithium diisopropylamide (LDA) to obtain oligomeric β-hydroxyl ester (3) in 87.2% yield; the repeating units of this oligomer were joined only by β-O-4 linkages as confirmed by nuclear magnetic resonance (NMR) spectroscopy. In the third step, the oligomeric β-hydroxyl ester (3) was reduced with LiAlH4 to give compound 4 in 42.4% yield. On the basis of NMR, matrix-assisted laser desorption ionization time-of-flight mass spectrometry, and gel
permeation chromatography analyses of compound 4, it was concluded that compound 4 was an oligomeric lignin model compound containing only β-O-4 interunit linkages. The number average degree of polymerization (DPn) of obtained compound 4 was about 7.0 (M
w/M
n = 1.42). Using this oligomeric lignin model compound, conventional degradation and analytical methods will give new information. 相似文献
19.
[目的]量化长白山原始阔叶红松林和杨桦次生林土壤颗粒有机碳和黑碳含量及分布特征,为研究东北森林土壤有机碳分布和积累提供依据。[方法]采用粒径分组方法测定原始阔叶红松林和杨桦次生林土壤表层(A_(11))和亚表层(A_(12))颗粒有机碳(POC)和黑碳(BC)含量,分析其与土壤基本性质(有机质、含水量、p H值、粘粒和团聚体等)的关系。[结果]表明:(1)两种森林类型土壤A_(11)和A_(12)POC含量分别为31.89 88.00、5.25 19.45 g·kg~(-1),BC含量分别为8.43 22.40、3.39 12.10 g·kg~(-1),二者随土壤深度增加而显著下降(p0.01)。(2)森林类型显著影响土壤POC和BC,表现为杨桦次生林原始阔叶红松林。(3)两种森林类型土壤POC与土壤有机质、含水量、水稳性团聚体均显著相关,与p H值、粘粒相关性不显著;BC与土壤p H值、粘粒和团聚体相关性均不显著,与含水量显著相关,与有机质仅在A_(11)显著相关。(4)土壤POC和BC显著相关(p0.01)。[结论]在长白山地区森林类型显著影响土壤POC和BC含量,杨桦次生林土壤POC和BC显著高于原始阔叶红松林,很大程度上与森林的采伐和火烧有关。两种森林类型土壤POC和BC分布格局是土壤物理化学性质综合作用的结果。 相似文献
20.
Michito Tsuyama Hajime Kobayashi Masaru Shinya Hisashi Yahata Yoshichika Kobayashi 《Journal of Forest Research》1996,1(2):79-85
Using attached and detached leaves ofAcer palmatum Thunb. andRhaphiolepsis umbellata Makino, pulse-modulated chlorophyll fluorescence and CO2 exchange were measured. Quantum yield of photosynthesis was determined from the fluorescence parameter(Fm′−Fs)/Fm′, where (Fm′−Fs) was defined as the difference between steady state chlorophyll fluorescence (Fs) and maximum fluorescence (Fm′) elicited by a saturating light pulse. The rate of electron transport through photosystem II (total electron flow) was calculated
from the product of quantum yield andA (PFD), whereA is the rate of absorbed photons as given by leaf absorptance, and PFD is the photon flux density at the leaf surface. The
rate of electron transport dependant on CO2 uptake (assimilative electron flow) was calculated from the gross photosynthetic rate in a leaf. The difference between the
rates of total and assimilative electron transport was denoted as the rate of non-assimilative electron transport which depends
on photorespiration and oxygen reduction. Available data provided quantitative information on the rate of non-assimilative
electron flow in intact leaves. When leaf photosynthesis ofA. palmatum was measured under sunlight, the rates of total and assimilative electron transport were determined to be approximately 900
and 150 μmol equiv. e/mg Chl·h, respectively. The difference (750 μmol equiv. e/mg Chl·h) was attributed to the activity of
non-assimilative electron flow. The ratio of total to assimilative electron flow was found to increase gradually with rising
in irradiance. The results suggest that non-assimilative electron flow occurred at much higher rate than assimilative electron
flow at high irradiance. Implications of the results are briefly discussed in relation to photosynthesis limitation in tree
leaves. 相似文献