木薯淀粉合成相关酶基因的克隆及表达分析

根据核苷酸同源性设计简并引物,克隆了一段大小为665 bp的木薯腺苷二磷酸葡萄糖焦磷酸化酶小亚基基因(sAGP)片段,并根据已知序列设计特异引物,克隆分离得到GBSS基因和SBE基因的cDNA片段.半定量RT-PCR分析结果表明,在木薯不同生长时期,3种淀粉合成相关酶基因的时空表达规律不同,GBSS基因表达有时期特异性和组织特异性,只在块根形成期和块根成熟期表达,在组织部位茎中表达最多,并且在这3种基因中表达水平相对较低.而sAGP基因在这3种基因中表达水平最高,在木薯决根成熟期竟达超量表达水平,并且在华南124和辐选0l中有品种差异,差异明显,在辐选01中的表达要优于华南124的表达.SBE基因表达较平稳,在各个时期表达水平都处于中间水平,在4个时期中,块根成熟期时的表达水平相对较高,块根形成期和苗期其次,收获期最低,也具有品种间差异,在辐选01中的表达要优于华南124的表达.     

木薯高、低粉品种贮藏根生长关键期的生理生化特性比较

通过研究木薯Manihot esculenta高粉和低粉品种贮藏根在发育关键期(形成–膨大期)主要生理生化性质的差异及其动态变化，为木薯栽培管理和育种改良提供理论基础。对高粉木薯品种‘辐选01’(FX01)和‘Kasetsart 50’(KU50)以及低粉品种‘华南124’(SC124)和‘9I’于种植后120、130、140、151和165 d五个时期分别取贮藏根样品，测定淀粉、可溶性糖、脱落酸(ABA)、保护酶、活性氧(ROS)、丙二醛(MDA)等生理生化指标并分析变化规律。结果表明，可溶性糖含量在贮藏根皮层中始终高于淀粉储藏区；高粉品种中总可溶性糖含量在贮藏根膨大期快速增加，且表现出较高的抗氧化酶活性和较低的ROS及MDA含量；ABA含量呈现出高粉品种比低粉品种低的趋势。木薯低粉和高粉品种中可溶性糖的分布与其转运后合成淀粉的规律相一致，高粉品种中淀粉的积累速率始终高于低粉品种，其较高的抗氧化酶活性和较低的ROS及MDA含量有利于木薯贮藏根中淀粉的积累。     

木薯块根淀粉高累积的相关酶的活性特征

木薯的价值在于其块根内高含量的淀粉。木薯淀粉不仅可以作为粮食,还可以作为原料制造乙醇燃料等。但是其块根中淀粉高效累积生化机制依然有待于解决。本研究比较性研究了大田生长的高、低淀粉木薯品种在4个生长关键时期的蔗糖和淀粉代谢相关的酶活性。结果表明,根中的淀粉合成途径的限速步骤酶腺苷二磷酸葡萄糖焦磷酸化酶(ADPGase)活性、叶部的蔗糖磷酸合成酶(SPS)活性与木薯块根内的淀粉含量呈正相关关系,而根淀粉降解酶活性则呈负相关关系。该研究表明通过提高根中的ADPGase和叶中的SPS活性,同时降低块根中的淀粉降解酶活性有可能可以提高木薯块根中的淀粉积累。     

施钾时期对食用甘薯光合特性和块根淀粉积累的影响

在施钾240 kg·hm^-2水平下,研究施钾时期对甘薯叶片光合特性和块根淀粉积累的影响.结果表明：与一次性施钾（全部基施）相比,分期施钾（1/2基施+1/2栽后75 d追施）提高了甘薯叶片的光合速率及叶片中磷酸蔗糖合成酶和块根中腺苷二磷酸焦磷酸化酶活性,提高了块根中淀粉积累速率(生育期平均增幅为6.7%),显著增加了块根产量(增幅为8.2%).与不施钾处理相比,两个施钾处理均提高了叶片中蔗糖的合成能力和蔗糖在块根中转化为淀粉的能力.     

木薯AGPase酶活性及其同工酶位点检测

本文以3个亲缘关系较远的高产木薯品种(SC124,SC8和Arg7)为材料,对块根发育过程中的AGPase活性和淀粉含量进行了测定,并利用AGPase同工酶电泳技术(非变性聚丙烯酰胺凝胶电泳)对AGPase酶活性较高时期同工酶位点进行检测,旨在分析木薯块根AGPase同工酶位点与AGPase酶活性和淀粉积累的关系.结果表明:木薯块根发育过程中AGPase酶活性和淀粉含量呈极显著正相关(R=0.802 12),Arg7的AGPase酶活性和淀粉含量在块根发育都显著高于SC124和SC8.同工酶分析表明,木薯至少有6个同工酶位点(AGPa,AGPb,AGPc,AGPd,AGPe和AGPf),且品种间具有多态性.其中,3个品种共有的4个同工酶位点分别为AGPa、AGPc、AGPd和AGPf,AGPe位点只在Arg7中出现,初步判断同工酶位点AGPe可能与其较高的酶活性和淀粉含量有关.本研究为系统研究木薯品种间AGPase酶活性与淀粉积累的关系提供了参考.     

摘心对‘巨峰’葡萄生长及蔗糖和淀粉代谢的调控作用

为揭示摘心对‘巨峰’葡萄(Vitis vinifera L. × V. labrusca L. cv. Kyoho)生长发育、蔗糖和淀粉代谢的作用及其分子机理,采用不同留叶摘心,研究‘巨峰’葡萄叶片和茎段表型,果实可溶性固形物、蔗糖和淀粉积累特征,以及蔗糖和淀粉代谢相关基因的表达变化。结果表明,摘心抑制‘巨峰’葡萄叶片生长和茎段增粗,但是促进花序早期的快速增长,提高单果重、果穗重、株产量和可溶性固形物含量;2叶摘心提高生长发育后期叶片蔗糖、茎段蔗糖和淀粉的含量;2叶摘心促进蔗糖代谢基因SPS、NI、CWI的高表达,同时抑制淀粉代谢中AMY的表达。因此,2叶摘心能够调控SPS、NI、CWI和AMY基因的表达进而促进蔗糖合成和淀粉积累,为果实成熟、新梢萌芽和开花奠定营养基础。     

木薯品种(系)类胡萝卜素代谢相关基因和蛋白表达水平分析

以白心木薯华南6068、华南9号、紫叶黄心木薯BGM019和粉红木薯Mirasol为材料,探究木薯块根膨大期和成熟期与类胡萝卜素代谢通路相关的14个基因和4种蛋白质表达水平变化。用HPLC检测块根β-胡萝卜素含量的变化,分别用qRT-PCR和Western blot方法对类胡萝卜素代谢通路相关基因和蛋白酶的表达水平进行分析。以华南6068为对照,研究结果表明:华南9号和紫叶黄心木薯BGM019成熟期中的类胡萝卜素合成途径关键基因PSY2、LCYB基因显著高于膨大期,而降解相关的关键基因CCD1、NCED3在成熟期的表达量显著低于膨大期(P0.05)。粉红木薯Mirasol成熟期中PSY2、LCYB的显著下调与CCD1、NCED3的显著上调(P0.05)是造成β-胡萝卜素含量差异的原因之一。通过分析不同木薯品种(系)在膨大期和成熟期块根类胡萝卜素代谢途径相关基因的表达水平,有助于解析β-胡萝卜素积累的分子机理。此外,Western blot结果显示抗坏血酸过氧化物酶、谷胱甘肽还原酶、超氧化物歧化酶和HSP70虽然和块根类胡萝卜素代谢途径没有直接关联,但它们在木薯膨大期和成熟期块根表达水平有显著差异(P0.05)。     

无机磷对叶片淀粉和蔗糖积累的影响

C_3植物大豆的叶片无机磷含量随年龄增长而降低,淀粉则显著增加。蔗糖积累数量较少,随叶龄增长略有增加。 不同浓度P_i处理的大豆植株,在光合不变的情况下,随着叶片中P_i含量的增加,淀粉积累不断减少,蔗糖不断增多。各种年龄的大豆叶片都在含较多P_i时有利于蔗糖合成,不利于淀粉合成。 C_4植物玉米经不同浓度P_i处理,叶淀粉与蔗糖的积累都稳定在一定水平上。P_i浓度过高时,积累数量都下降。在C_4植物玉米叶组织中,P_i含量的变化不是调节淀粉与庶糖合成的重要因素。在C_3植物大豆和C_4植物玉米的叶组织中,P_i对淀粉和蔗糖合成的影响是不同的。     

南荻纤维素合成的相关生理特性

为探明南获纤维素合成相关的生理特性,研究了盆栽条件下南荻生长过程中IAA、ABA含量和纤维素合成关键酶（蔗糖合成酶、β-1,3-葡聚糖酶）活性的变化及其与纤维素含量间的关系。结果显示,在整个生育期,叶片中IAA含量、蔗糖合成酶活性和β-1,3-葡聚糖酶活性变化趋势一致,均呈先升高后降低的单峰曲线变化;叶片中ABA含量与茎秆蔗糖含量均先降低后增加再降低;茎秆纤维素、半纤维素和木质素含量均随生育进程呈现升高趋势。生育前期为调控纤维素合成的关键时期,此时ABA／IAA比值下降,蔗糖合成酶和β-1,3-葡聚糖酶活性快速上升,有利于纤维素的快速积累。     

叶片光合产物输出的抑制与淀粉和蔗糖的积累

选择糖叶、淀粉叶及中间型植物叶片用水烫法处理叶片基部或将离体叶片插在水中,或用环剥的方法阻碍叶片输出光合产物与自然对照对比分析,观察输出与否对叶片淀粉与蔗糖合成的影响。 糖叶与中间型叶片在进行光合作用的同时有相当数量的光合产物输出,淀粉叶在光合作用的同时很少光合产物输出。糖叶输出停滞时主要增加蔗糖的积累。中间型叶片输出停滞时蔗糖与淀粉的积累均比对照增加。淀粉叶处理与否同淀粉与蔗糖的积累无显著差异。     

HPLC-ELSD法分析木薯韧皮部汁液主要糖成分

采用高效液相色谱—蒸发光散射检测法(HPLC-ELSD)对木薯韧皮部汁液糖成分进行了分析.结果表明:与大多数木本植物一样,其同化物的主要运输形式是蔗糖,并未发现糖醇类和棉子糖等寡糖.对块根产量、淀粉含量较低的半野生种W14的对比实验发现,无论是蔗糖还是己糖含量都大大地低于栽培种,说明蔗糖是木薯块根淀粉累积的主要来源,对其累积速率和量起着决定性作用.结果同时证明此方法可以高效、快速简便地定性和定量测定木薯韧皮部汁液中的糖类.     

半干旱雨养农业区马铃薯干物质和钾素积累与分配特性

2010年在甘肃省定西市通过大田试验,研究了半干旱雨养农业区马铃薯的干物质(DM)和钾素(K)积累与分配规律.结果表明： 马铃薯根、茎、叶的DM积累量在全生育期内呈单峰曲线,大小顺序为叶＞茎＞根;全株和块茎DM积累量在全生育期内持续增加,均呈“S”型增长曲线.全株的DM积累最大速率大于块茎,且出现时间比块茎早17 d. DM在各器官中的分配以块茎形成末期和块茎增大末期为转折点;在块茎形成末期之前,DM分配比例以叶片最大,其后以块茎最大;干质量平衡期出现在块茎增大末期(出苗后90 d左右),此期之前块茎的DM积累量小于根茎叶,二者的积累量呈正相关,其后块茎大于根茎叶,二者呈负相关.全株干物质积累主要来源于块茎.植株干物质积累量在苗期、块茎形成期、块茎增长期、淀粉积累期和成熟期的分配比例分别为5%、30%、60%、4%和1%,块茎干物质积累量的分配比例分别为0、18%、62%、18%和2%.全生育期内,马铃薯50%以上的干物质在块茎增长期形成. 马铃薯品种“新大坪”各器官中钾浓度以茎最高,块茎最低,在干质量平衡期之前根中的钾浓度高于叶,之后低于叶;各器官中钾的积累量在干质量平衡期之前集中分配在根茎叶中,表现为茎＞叶＞根,之后主要向块茎中分配和积累,至成熟期,60%以上的钾贮藏在块茎中.     

Effect of Day Length and Night Temperature on Starch Accumulation and Degradation in Soybean

The effect of the day length on the accumulation and the degradationof the starch in leaf, stem and root tissues of prefloweringsoybean plants was determined by growing plants under a 7 or14 h light regime. As has been reported previously, the rateof starch accumulation by leaves was inversely related to daylength. High sucrose content was associated with a high rateof starch accumulation. Stem tissue showed diurnal fluctuationsin starch content and the rate of accumulation was also inverselyrelated to day length. This starch resulted from photosynthesiswithin the stem itself. A negligible amount of starch was foundin root tissue of both sets of plants. The rate of starch breakdown in leaves of 7 h plants was significantlyless than that in 14 h plants. Nevertheless, leaf starch inshort day length plants was depleted at least 4 h prior to theend of the dark period. In both sets of plants, degradationof stem starch started simultaneously with that in the leavesand continued throughout the dark period, although at a muchlower rate than that of leaves. Thus, stem starch acted as abuffer once leaf starch was depleted, providing carbohydratesto the plant, although in small quantities. To determine if soybean leaves adjust their rate of starch accumulationduring the light period to different dark period temperatures,plants were grown under temperature regimes of 30/20 ^°Cand 30/30 ^°C. Plants did not differ in rate of starch accumulationor CO₂ exchange rate, but did show large differences in growthcharacteristics. High temperature plants had significantly greaterleaf area and tended to have greater leaf area ratio. Thus,despite similar rates of starch accumulation on a leaf areabasis, high temperature plants accumulated greater amounts ofstarch on a per plant basis. Glycine max(L.)Merr., soybean reserve carbohydrates, remobilization, source-sink realtionships     

An ordered EST catalogue and gene expression profiles of cassava (<Emphasis Type="Italic">Manihot esculenta</Emphasis>) at key growth stages

A cDNA library was constructed from the root tissues of cassava variety Huanan 124 at the root bulking stage. A total of 9,600 cDNA clones from the library were sequenced with single-pass from the 5′-terminus to establish a catalogue of expressed sequence tags (ESTs). Assembly of the resulting EST sequences resulted in 2,878 putative unigenes. Blastn analysis showed that 62.6% of the unigenes matched with known cassava ESTs and the rest had no ‘hits’ against the cassava database in the integrative PlantGDB database. Blastx analysis showed that 1,715 (59.59%) of the unigenes matched with one or more GenBank protein entries and 1,163 (40.41%) had no ‘hits’. A cDNA microarray with 2,878 unigenes was developed and used to analyze gene expression profiling of Huanan 124 at key growth stages including seedling, formation of root system, root bulking, and starch maturity. Array data analysis revealed that (1) the higher ratio of up-regulated ribosome-related genes was accompanied by a high ratio of up-regulated ubiquitin, proteasome-related and protease genes in cassava roots; (2) starch formation and degradation simultaneously occur at the early stages of root development but starch degradation is declined partially due to decrease in UDP-glucose dehydrogenase activity with root maturity; (3) starch may also be synthesized in situ in roots; (4) starch synthesis, translocation, and accumulation are also associated probably with signaling pathways that parallel Wnt, LAM, TCS and ErbB signaling pathways in animals; (5) constitutive expression of stress-responsive genes may be due to the adaptation of cassava to harsh environments during long-term evolution.     

The contribution of plastidial phosphoglucomutase to the control of starch synthesis within the potato tuber

The aim of this work was to evaluate the extent to which plastidial phosphoglucomutase (PGM) activity controls starch synthesis within potato (Solanum tuberosum L. cv. Desirée) tubers. The reduction in the activity of plastidial PGM led to both a correlative reduction in starch accumulation and an increased sucrose accumulation. The control coefficient of plastidial PGM on the accumulation of starch was estimated to approximate 0.24. The fluxes of carbohydrate metabolism were measured by investigating the metabolism of [U-14C]glucose in tuber discs from wild-type and transgenic plants. In tuber discs the control coefficient of plastidial PGM over starch synthesis was estimated as 0.36, indicating that this enzyme exerts considerable control over starch synthesis within the potato tuber.     

Evidence of the crucial role of sucrose synthase for sink strength using transgenic potato plants (Solanum tuberosum L.)

Sink strength of growing potato tubers is believed to be limited by sucrose metabolism and/or starch synthesis. Sucrose synthase (Susy) is most likely responsible for the entire sucrose cleavage in sink tubers, rather than invertases. To investigate the unique role of sucrose synthase with respect to sucrose metabolism and sink strength in growing potato tubers, transgenic potato plants were created expressing Susy antisense RNA corresponding to the T-type sucrose synthase isoform. Although the constitutive 35S CaMV promotor was used to drive the expression of the antisense RNA the inhibition of Susy activity was tuber-specific, indicating that independent Susy isoforms are responsible for Susy activity in different potato organs. The inhibition of Susy leads to no change in sucrose content, a strong accumulation of reducing sugars and an inhibition of starch accumulation in developing potato tubers. The increase in hexoses is paralleled by a 40-fold increase in invertase activities but no considerable changes in hexokinase activities. The reduction in starch accumulation is not due to an inhibition of the major starch biosynthetic enzymes. The changes in carbohydrate accumulation are accompanied by a decrease in total tuber dry weight and a reduction of soluble tuber proteins. The reduced protein accumulation is mainly due to a decrease in the major storage proteins patatin, the 22 kDa proteins and the proteinase inhibitors. The lowered accumulation of storage proteins is not a consequence of the availability of the free amino acid pool in potato tubers. Altogether these data are in agreement with the assumption that sucrose synthase is the major determinant of potato tuber sink strength. Contradictory to the hypothesis that the sink strength of growing potato tubers is inversely correlated with the tuber number per plant, no increase in tuber number per plant was found in Susy antisense plants.     

Control of Photosynthetic Sucrose Synthesis by Fructose 2,6-Bisphosphate : II. Partitioning between Sucrose and Starch

The role of fructose 2,6 bisphosphate in partitioning of photosynthate between sucrose and starch has been studied in spinach (Spinacia oleracea U.S. hybrid 424). Spinach leaf material was pretreated to alter the sucrose content, so that the rate of starch synthesis could be varied. The level of fructose 2,6-bisphosphate and other metabolites was then related to the accumulation of sucrose and the rate of starch synthesis. The results show that fructose 2,6-bisphosphate is involved in a sequence of events which provide a fine control of sucrose synthesis so that more photosynthate is diverted into starch in conditions when sucrose has accumulated to high levels in the leaf tissue. (a) As sucrose levels in the leaf rise, there is an accumulation of triose phosphates and hexose phosphates, implying an inhibition of sucrose phosphate synthase and cytosolic fructose 1,6-bisphosphatase. (b) In these conditions, fructose 2,6-bisphosphate increases. (c) The increased fructose 2,6-bisphosphate can be accounted for by the increased fructose 6-phosphate in the leaf. (d) Fructose 2,6-bisphosphate inhibits the cytosolic fructose 1,6-bisphosphatase so more photosynthate is retained in the chloroplast, and converted to starch.     

不同光照条件下三七幼苗形态及生长指标的变化

用遮阳网设置不同透光率(自然全光照的1%、3%、8%、12%和22%)处理,对不同光照条件下三七〔Panax notoginseng(Burk.)F.H.Chen〕幼苗形态指标(株高、冠幅、块根长、主根长、块根直径、茎基径、单株须根数和单株须根长)、干物质积累(不同器官干质量)和分配以及叶片性状(单株叶面积、比叶面积和叶绿素相对含量SPAD值)的变化进行了研究。结果表明:在透光率不同的条件下三七幼苗的形态指标、不同器官干质量及分配以及叶片性状均有明显变化。其中,块根直径、单株须根长、单株须根数、不同器官(块根、须根、根、叶片和茎)干质量和植株总干质量均随透光率增大逐渐提高;株高在透光率22%条件下最高;冠幅和单株叶面积在透光率3%条件下最大;主根长、茎基径、根冠比、根质比及SPAD值均在透光率8%条件下最高;茎质比和叶质比在透光率3%和1%条件下较大;比叶面积随透光率增大逐渐降低。综合分析结果揭示:三七是一种典型的喜阴植物,种植过程中适当遮阳有利于其生长和干物质积累,其中透光率8%对三七幼苗生长较为适宜。