Evaluation of Root Sink Ability of Sweet Potato (Ipomoea batatas Lam.) Cultivars on the Basis of Enzymatic Activity in the Starch Synthesis Pathway

Activities of five enzymes, sucrose synthase (SUS), uridine 5'-diphosphate glucose pyrophosphorylase (UDPGPPase), fructose-1,6 bisphosphatase (FBPase), adenosine 5'diphosphate glucose pyrophosphorylase (ADPGPPase) and starch synthase (STS), in the metabolic pathway of starch synthesis, were compared between two sweet potato ( Ipomoea batatas Lam.) cultivars, Koganesengan (C.V.KOG, a recently released high yield cultivar) and Tsurunasigenji (C.V.TSU, an old, local cultivar with poor yield). The measurements were carried out using the root samples (tuberous, thick and fibrous roots) harvested at the fast tubering stage. Of the five enzymes, SUS, ADPGPPase and STS showed high activities in the tuberous root, particularly in that of C.V.KOG, and a similar trend was observed for activities of these enzymes on a protein basis. The increased activity of the three enzymes is considered to be one of the characteristics in a high yield cultivar, allowing the root to function effectively as a starch synthesis and storage organ.     

Astragalus polysaccharides protects against free fatty acid-induced human vascular endothelial cell dysfunction via AMPK-eNOS pathway

AIM: To study the protective effect of Astragalus polysaccharides (APS) on free fatty acid-induced injury in human umbilical vein endothelial cells (HUVECs). METHODS: Cultured HUVECs were divided into control group, APS group [APS (200 mg/L) treated for 24 h], free fatty acid group [free fatty acid (0.25 mmol/L) treated for 24 h], free fatty acid plus APS group [free fatty acid (0.25 mmol/L) and APS (200 mg/L) treated for 24 h], and compound C group [free fatty acid (0.25 mmol/L) and APS (200 mg/L) and AMPK inhibitor compound C (10 μmol/L) treated for 24 h]. The cell viability was detected by MTT assay. Nitric oxide (NO) content in the medium was determined by nitrate reductase assay. The protein levels of total adenosine monophosphate-activated protein kinase (AMPK), phosphorylated adenosine monophosphate-activated protein kinase (p-AMPK), endothelial nitric oxide synthase (eNOS) and phosphorylated endothelial nitric oxide synthase (p-eNOS) were measured by Western blot. RESULTS: No significant difference of all indexes between APS group and control group was observed. The cell viability in free fatty acid group decreased significantly compared with control group. The cell viability in free fatty acid plus APS group was significantly improved as compared with free fatty acid group. The cell viability in compound C group was almost the same as that in free fatty acid group. The No content and protein levels of p-AMPK and p-eNOS in free fatty acid group decreased obviously as compared with control group, while the NO content and protein levels of p-AMPK and p-eNOS in free fatty acid plus APS group increased obviously compared with free fatty acid group. No significant difference of the p-AMPK and p-eNOS protein levels between free fatty acid plus APS group and free fatty acid group was observed. No significant difference of the AMPK and eNOS protein levels in all groups was found. CONCLUSION: APS attenuates the free fatty acid-induced injury, and its mechanism is related to the AMPK-eNOS signal pathway.     

Effects of eplerenone on expression and activity of aortic endothelial nitric oxide synthase in hypertensive rats induced by high-salt intake

AIM: To explore the effects of eplerenone on the expression and activity of aortic endothelial nitric oxide synthase(eNOS) in high salt-induced hypertensive rats.METHODS: Male Wistar rats(4 week old, weighting 50～60 g) were randomly divided into control group, high-salt diet group and eplerenone group. The rats in control group were fed with ordinary rodent animal diet, the rats in high-salt group and eplerenone group were exposed to 5% salt diet for 16 weeks and administrated with the same dosage of saline or eplerenone(40 mg·kg^-1·d^-1) by gavage for 4 weeks, respectively. Systolic blood pressure(SBP) was measured by tail-cuff every 2 weeks. The rats were sacrificed after 16 weeks and the thoracic aorta was collected. The aldosterone content in the aorta was measured by ELISA. The protein levels of mineralocorticoid receptor(MR) and eNOS were determined by Western blot. The activitie of constitutive NOS(cNOS) was measured by chemocolorimetry. The protein localization of eNOS, neuronal nitric oxide synthase(nNOS) and MR was observed by immunohistochemistry.RESULTS: A process of 8-week high-salt diet increased SBP gradually. SBP in the rats exposure to high salt for 16 weeks was significantly higher than that in control group(P<0.05). After 4 weeks of eplerenone treatment, SBP in the rats was significantly lower than that before treatment(P<0.05). Compared with control group, the aldosterone content in the aorta were significantly increased in high-salt diet group and eplerenone group(P<0.05), the expression level of MR also increased significantly(P<0.05). Compared with control group, both eNOS protein expression(P<0.05) and cNOS activity in high-salt diet group were significantly decreased(P<0.05). The protein expression of eNOS as well as cNOS activity in aorta increased significantly in eplerenone group compared with high-salt diet group(P<0.05).CONCLUSION: Aldosterone content in aorta of high-salt-induced hypertensive rats increases significantly. Aldosterone attenuates the protein expression of eNOS and reduces the enzyme activity through the activation of mineralocorticoid receptor. The selective mineralocorticoid receptor antagonist eplerenone enhances the protein expression of eNOS and its activity, thereby improves eNOS function.     

水稻灌浆期籽粒中3个与淀粉合成有关的酶活性变化

在水培和盆栽条件下，研究了6个水稻品种(含籼/粳杂交组合、新株型品系)灌浆期强、弱势粒中ADPG焦磷酸酶(EC2.7.7.21)、淀粉合成酶(EC2.4.1.21)和淀粉分枝酶或Q-酶(EC2.4.1.18)的活性变化及其与灌浆充实的关系。3个酶的活性变化与籽粒灌浆动态相关联：淀粉酶最高活性出现的时间稍前或同步于最大灌浆速率的时间；ADPG焦磷酸酶     

Characterization of waxy grain sorghum lines in relation to granule-bound starch synthase

The waxy phenotype, associated with endosperm containing little or no amylose, has been recognized in sorghum (Sorghum bicolor L. Moench) since 1933. Although variants of the waxy gene are well characterized in other cereals, the waxy trait has been assumed to be controlled by a single allele, wx, in sorghum. Recent improvements in technologies encourage re-examination of the waxy sorghums. The objectives of this research were therefore to identify and characterize sorghum lines with differing waxy alleles and to describe the actions of those alleles in crosses. Grain of eight waxy sorghum lines (BTxARG1, BTx630, Tx2907, B.9307, 94C274, 94C278, 94C289, 94C369), three wild-type checks (BWheatland, RTx430, BN122), and F₂ families from crosses among a subset of these lines were evaluated for presence or absence of granule-bound starch synthase (GBSS), the gene product of the wx locus, and wild-type vs. waxy endosperm. The F₂ segregation ratios were tested for fit to a 3:1 ratio using Chi-square analyses. Two distinctly different naturally occurring waxy alleles were identified: One with no GBSS (GBSS−), and one with apparently inactive GBSS present (GBSS+). We propose that the waxy allele with no GBSS be designated wx^a, and that waxy allele with apparently inactive GBSS present be designated wx^b. These two alleles are located in close proximity on the waxy locus. The wx^b allele is dominant to the wx^a allele in terms of GBSS production, and both are recessive to the wild-type Wx in terms of amylose content. The U.S. Government's right to retain a non-exclusive, royalty-free license in and to any copyright is acknowledged.     

环境胁迫和激素诱导甘蔗ACC合成酶基因家族三个成员的表达

ACC合成酶（ACS）是高等植物乙烯生物合成途径中的限速酶。在克隆到甘蔗ACC合成酶基因家族3个成员Sc-ACS1、Sc-ACS2和Sc-ACS3的基础上，分别以它们作为探针，检测了激素诱导和环境胁迫对甘蔗苗期该3成员表达的影响。结果表明，乙烯利诱导Sc-ACS1在叶中表达，在根中不表达；Sc-ACS2在根、叶中表达；Sc-ACS3主要在根部表达，在叶中不表达。在甘蔗叶片中，Sc-ACS1对冷胁迫、暗培养和LiCl胁迫均有应答，并受植物生长激素IAA诱导而表达；Sc-ACS2无论是在生长激素诱导还是环境胁迫下，其mRNA均维持较低水平。     

Field evaluation of antisense RNA mediated inhibition of GBSS gene expression in potato

Summary Granule-bound starch synthase (GBSS) catalyses the synthesis of amylose in starch granules. Analysis of antisense RNA mediated inhibition of GBSS gene expression in large numbers of tubers from in vitro grown, greenhouse grown and field grown transgenic potato plants revealed stable and total inhibition of GBSS gene expression in one clone. In three other transgenic genotypes partial and unstable inhibition was found. In these genotypes both GBSS activity and amylose content were remarkably reduced compared with the non-transformed control genotype. No relationship was found between the level of inhibition of GBSS gene expression and yield and dry matter content.     

Developing gene-tagged molecular markers for functional analysis of starch-synthesizing genes in rice (Oryza sativa L.)

The granule-bound starch synthase(GBSS), starch branching enzymes 1 (SBE1)and 3 (SBE3) are major enzymes involved in starch biosynthesis in rice endosperm. Available sequences of Sbe1 and Sbe3 genes encoding corresponding SBE1 and SBE3 have been used to identify homologous regions from genome databases of both the indica rice 93-11 and the japonica rice Nipponbare. Sequence diversities were exploited to develop gene-tagged markers to distinguish an indica allele from a japonica allele for both Sbe1 and Sbe3 loci. With these newly developed gene-tagged markers and available Wx gene markers, the roles of these starch-synthesizing genes (Sbe1, Sbe3, and Wx) in determining amylose content (AC) in the rice endosperm were evaluated using a double-haploid (DH)population derived from a cross between the indica rice cv. Nanjing11 and the japonica rice cv. Balilla. Only the Wx and Sbe3 loci had significant effects on the AC variation. On average, indica Wx ^a genotypes showed ∼9.1% higher AC than japonica Wx ^b genotypes, while indica Sbe3 ^a genotypes showed ∼1.0% lower AC than japonica Sbe3 ^b genotypes. A significant interaction was also observed between Wx and Sbe3 loci whereby the amylose content was 0.3% higher in Sbe3 ^a than Sbe3 ^b genotypes in the presence of the Wx ^a allele, but it was lower by 2.3% in the presence of the Wx ^b allele. Overall, polymorphisms at the Wx and Sbe3 loci together could account for 79.1% of the observed AC variation in the DH population. The use of gene-tagged markers in marker-assisted selection and gene functional analysis was also discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

旱种水稻结实期茎中碳同化物的运转及其生理机制

以杂交水稻汕优63和籼稻扬稻6号为材料，研究了旱种(地膜覆盖栽培)水稻结实期茎中碳同化物的运转及其生理机制。结果表明，抽穗至成熟期旱种水稻标记14C从茎中向籽粒的再分配、茎中非结构性碳水化合物(NSC)的运转率及其对籽粒的贡献率均显著高于水种(常规栽培)水稻。水稻旱种后籽粒中的蔗糖合成酶和酸性转化酶活性、茎中α-淀