Comparison of methods for protein extraction from pine needles

Extraction of proteins from pine needles for proteomic analysis has long been a challenge for scientists. We compared three different protein extraction methods including sucrose, Tris-HCl and trichloroacetic acid （TCA）/acetone （TCA method） to determine their efficiency in separating pine needle proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis （SDS-PAGE） and two-dimensional PAGE （2D-PAGE）. Proteins were then separated by SDS-PAGE. Among three methods the method using sucrose extraction buffer showed the highest efficiency and highest quality in separating proteins. In addition, clearer and more stable strips were detected by SDS-PAGE using sucrose extraction buffer. When the proteins extracted using sucrose extraction buffer were separated by 2D-PAGE, more than 300 protein spots, with isoelectric points （PI） ranging from 4.0 to 7,0 and molecular weights （MW） from 6.5 to 97.4 kD, were observed. This confirmed that the method with sucrose extraction buffer was an efficient and reliable method for extracting proteins from pine needles.     

蔗糖酯的特性及在食品中的应用

蔗糖酯是一种安全的食品乳化剂,有宽范围的HLB值,具有优良的物理化学性质,在食品中应用广泛。     

蔗糖浓度对葡萄试管苗生长及光合作用的影响

糖为植物生长发育提供能量,也是形态建成的物质基础。但培养基的蔗糖浓度对葡萄试管苗生长及光合作用的影响研究较少,以‘黑比诺’葡萄试管苗为试材,以GS为基本培养基、蔗糖浓度30g/L为对照,分别研究蔗糖浓度0、60g/L对‘黑比诺’葡萄试管苗生理指标和光合参数的影响。结果表明:蔗糖浓度为30g/L试管苗生长状况最佳,试管苗干鲜重、叶绿素含量、可溶性糖含量均显著高于其他2个处理,总糖含量无显著差异;蔗糖浓度60 g/L处理试管苗叶片可溶性蛋白含量显著高于其他2个处理,在高糖及无糖处理下,试管苗叶片蒸腾速率(E)、气孔导度(Gs)与净光合速率(Pn)均显著低于正常浓度处理;叶绿素a、叶绿素b、光合电子传递速率(ETR)、最大量子产量(Fv/Fm)在蔗糖浓度30 g/L处理下均显著高于其他2个处理,叶片中不同激素受蔗糖浓度的影响各不相同。综上,蔗糖浓度能够影响葡萄试管苗生物量、叶绿素相对含量、可溶性蛋白、可溶性糖及总糖含量变化,培养基蔗糖浓度过高或过低会影响叶绿素荧光参数的变化,进而影响光合作用。     

番茄果实生长发育过程中糖的代谢

通过测定番茄果实生长发育过程中蔗糖、果糖、可溶性总糖、淀粉的含量和糖代谢中涉及到的蔗糖合成酶、蔗糖磷酸合成酶、酸性转化酶、中性转化酶的活性,研究了番茄果实生长发育过程中糖的代谢过程。根据番茄果实的生长发育曲线和相对日生长率,可以将番茄的生长发育过程按开花后39 d作为分界点分为2个阶段,39 d之前为产量形成期,之后为品质形成期。在产量形成期内,果糖和可溶性总糖含量逐渐升高,淀粉含量逐渐下降,蔗糖含量先升高后下降,变化不剧烈,以积累淀粉为主。同期AI和NI的活性较低,SS和SPS的活性较高。在品质形成期,果糖含量升高,淀粉含量下降,蔗糖和可溶性总糖变化不明显,以积累果糖为主。AI和NI的活性急剧升高,SS和SPS的活性很低。统计分析表明,番茄果实单果重量与淀粉含量、淀粉含量与NI活性呈显著负相关,可以参考淀粉的含量和NI的活性进行番茄高产研究。果糖含量与淀粉含量呈显著负相关,与SS和AI,NI的活性均为显著正相关;番茄果实中的可溶性总糖与淀粉含量呈显著负相关,与SS的活性呈显著正相关,可以参考SS和AI,NI的活性进行高果糖含量的品质研究。     

Genotypic Variation in Wheat in Response to Water Stress and Abscisic Acid-Induced Accumulation of Osmolytes in Developing Grains

Two wheat genotypes differing in water stress sensitivity (C306, relatively tolerant; HD2329, relatively susceptible) were water stressed during early grain filling by withholding water for 7 days at watery‐ripe (WR) stage and examined for water status, abscisic acid (ABA) and osmolytes in grains as well as in flag leaf (FL). Both the genotypes differed significantly from each other in endogenous levels of ABA, proline, glycine betaine, total soluble sugars, reducing sugars, sucrose and potassium. The tolerant genotype showed higher content of ABA, proline, glycine betaine, total sugars, reducing sugars and had higher water content in its FL and grains than the susceptible genotype, which contained more of glycine beatine and potassium but had lower ABA and water content in its FL and grains. Although carbohydrates constituted the major amount of all the solutes, proline and glycine betaine increased manifold during stress. Exogenous application of 2 μm ABA at 5 days after anthesis to FL of stressed plants increased the endogenous content of ABA, accelerated the accumulation of osmolytes, improved the water status of FL and grains that resulted in higher grain weight, especially in the susceptible genotype. Differential response of contrasting wheat genotypes to water stress may be governed by ABA‐dependent solute accumulation in grains and FL.     

毛白杨蔗糖合酶基因PtSUS1的克隆、表达及单核苷酸多态性分析

利用RT-PCR方法,首次从毛白杨成熟木质部cDNA文库中分离出PtSUS1 cDNA全长,并进行了测序和序列分析。结果表明:克隆的毛白杨PtSUS1 cDNA片段总长为2703bp,基因内部含有完整的开放阅读框架,大小为2415bp,可编码长度为805个氨基酸残基的蛋白质,所推导的蛋白质氨基酸序列与拟南芥AtSUS1、水稻OsSUS1的蛋白质氨基酸序列同源性分别为83.4%,75.7%。组织特异性Realtime-PCR结果显示,PtSUS1在成熟木质部表达丰度最高,在成熟叶片、根部和正在发育的木质部表达丰度中等,在韧皮部和形成层有少量表达,在嫩叶与顶端分生组织中表达丰度最低。在此基础上,组合利用MEGA4.0和DnaSP4.50.4软件对毛白杨40株基因型个体的PtSUS1序列进行比对和分析,共检测到235个单核苷酸多态性(SNP)位点,SNP频率为1/25bp,多样性指数π为0.00924。在外显子区域,共检测到66个SNP位点,其中55个为同义突变,10个为错义突变,1个为无义突变。在编码区内,非同义突变与同义突变的比率<1,这一结果显示在毛白杨物种演化过程中,纯化选择是PtSUS1基因内同义SNP位...     

杉木幼苗叶片蔗糖和淀粉含量的昼夜变化

杉木幼苗叶片蔗糖和淀粉含量的昼夜变化＊苏梦云周国璋金正法关键词杉木、蔗糖、淀粉、昼夜变化杉木（Ｃｕｎｎｉｎｇｈａｍｉａｌａｎｃｅｏｌａｔａ（Ｌａｍｂ．）Ｈｏｏｋ．）是我国南方重要造林树种，不同无性系间的生长存在明显差异。从光合作用方面探讨杉木速生生理...     

Function Analysis of Sucrose Transporter OsSUT4 in Sucrose Transport in Rice

【Objective】The transmembrane transport of sucrose is mainly mediated by sucrose transporters, among which five SUT coding genes have been identified in rice. However, the role of OsSUT4 in the sucrose transport is unclear. 【Method】CRISPR/Cas9 techniques were used to construct ossut4-deleted mutants. The ossut4 mutant was featured by lower plant height and grain yield as well as higher tiller number as compared with wild type plants.【Result】Sucrose and starch accumulation in the leaf of the ossut4 mutants lead to slower net photosynthetic rate; insufficient sugar supply results in the postpone of grain filling. Base on the subcellular and histochemical localization, OsSUT4 was located on the cytoplasmic membrane. Tissue localization showed that OsSUT4 was expressed in the embryo of germinating seed, vascular bundles of coleoptile, glume shell, anthers at flowering stage and aleurone layer of caryopsis at filling stage. 【Conclusion】OsSUT4 plays an important role in sucrose transport.     

Effects of long‐term elevation of air temperature on sucrose metabolism in cotton leaves at different positions

Global warming could possibly increase the air temperature by 1.8–4.0°C in the coming decade. To forecast the effects of long‐term elevation of air temperature on sucrose synthesis in cotton leaves at different positions, field experiments with the treatments of ambient temperature (control) and elevated temperature (2–3°C) across cotton's boll forming stage were conducted in 2011 and 2012. Results showed that sucrose content in the fourth main‐stem leaves from the terminal (FMLT) during 10–17 days after treatment (DAT) and in the leaves subtending the cotton boll (LSCB) during 17–45 days post‐anthesis (DPA) was lower in the increased temperature treatment than control treatment, but starch content was higher, resulting in higher specific leaf weight (SLW). Lower sucrose content might be attributed to low ribulose‐1,5‐bisphosphate carboxylase‐oxygenase (Rubisco) and cytosolic fructose‐1,6‐bisphosphatase (cy‐FBPase) activities under elevated temperature for all tested leaves. Comparison of different fruiting branch (FB) positions, sucrose transformation rate just decreased in LSCB growing on FB_2–3 and FB_6–7. Compared with FB_10–11, the leaves at FB_2–3 and FB_6–7 were more sensitive to the increased temperature, showing as that the changing amplitudes of SLW, sucrose content, starch content, initial Rubisco activity and cy‐FBPase activity were greater. Regarding different types of cotton leaves, the effects of elevated temperature on sucrose content, starch content and initial Rubisco activity in FMLT were obvious at the early stage (10–17 DAT), but were not obvious at the late stage. However, the effects were consistent in LSCB from 17 to 45 DPA. In addition, long‐term elevated temperature had smaller impacts on above indices in FMLT than LSCB. Sucrose phosphate synthase (SPS) and sucrose synthase (SuSy) activities were decreased by elevated temperature in FMLT at the early stage, but were increased in LSCB, suggesting that SPS and SuSy were the key enzymes between FMLT and LSCB in response to elevated temperature.     

有机无机肥配施对水稻光合特性及NR与SPS活性的影响

采用田间小区试验,研究了有机肥(稻草、猪粪、猪粪堆肥和沼渣沼液)与化肥配施对水稻光合特性、硝酸还原酶(NR)及蔗糖磷酸合成酶(SPS)活性的影响.结果表明,猪粪堆肥与化肥配施处理在提高水稻功能叶净光合速率、水稻碳氮代谢关键时期NR和SPS酶活性方面较其他类型的配施处理有较明显优势,并能提高水稻产量和改善品质.与施纯化肥...