Identification and characterization of granule bound starch synthase I (GBSSI) gene of tartary buckwheat (Fagopyrum tataricum Gaertn.)

Tartary buckwheat (Fagopyrum tataricum Gaertn.) is increasingly considered as an important functional food material because of its rich nutraceutical compounds. Reserve starch is the major component of tartary buckwheat seed. However, the gene sequences and the molecular mechanism of tartary buckwheat starch synthesis are unknown so far. In this study, the complete genomic sequence and full-size cDNA coding tartary buckwheat granule-bound starch synthase I (FtGBSSI), which is responsible for amylose synthesis, were isolated and analyzed. The genomic sequence of the FtGBSSI contained 3947 nucleotides and was composed of 14 exons and 13 introns. The cDNA coding sequence of FtGBSSI shared 63.3%–75.1% identities with those of dicots and 56.6%–57.5% identities with monocots (Poaceae). In deduced amino acid sequence of FtGBSSI, eight motifs conserved among plant starch synthases were identified. A cleavage at the site IVC↓G of FtGBSSI protein produces the chloroplast transit sequence of 78 amino acids and the mature protein of 527 amino acids. The FtGBSSI mature protein showed an identity of 73.4%–77.8% with dicot plants, and 67.6%–70.4% with monocot plants (Poaceae). The mature protein was composed of 20 α-helixes and 16 β-strands, and folds into two main domains, N- and C-terminal domains. The critical residues which are involved in ADP and sugar binding were predicted. These results will be useful to modulate starch composition of buckwheat kernels with the aim to produce novel improved varieties in future breeding programs.     

Ectopic expression of FaesAP3, a Fagopyrum esculentum (Polygonaceae) AP3 orthologous gene rescues stamen development in an Arabidopsis ap3 mutant

Arabidopsis thaliana APETALA3 (AP3) and Antirrhinum majus DEFICIENS (DEF) MADS box genes are required to specify petal and stamen identity. AP3 and DEF are members of the euAP3 lineage, which arose by gene duplication coincident with radiation of the core eudicots. In order to investigate the molecular mechanisms underlying organ development in early diverging clades of core eudicots, we isolated and identified an AP3 homolog, FaesAP3, from Fagopyrum esculentum (buckwheat, Polygonaceae), a multi-food-use pseudocereal with healing benefits. Protein sequence alignment and phylogenetic analyses revealed that FaesAP3 grouped into the euAP3 lineage. Expression analysis showed that FaesAP3 was transcribed only in developing stamens, and differed from AP3 and DEF, which expressed in developing petals and stamens. Moreover, ectopic expression of FaesAP3 rescued stamen development without complementation of petal development in an Arabidopsis ap3 mutant. Our results suggest that FaesAP3 is involved in the development of stamens in buckwheat. These results also suggest that FaesAP3 holds some potential for biotechnical engineering to create a male sterile line of F. esculentum.     

普通荞麦资源的耐铝性研究

利用小容器溶液培养法对耐铝性鉴定条件和52份普通荞麦栽培品种资源的耐铝性进行了研究。结果发现普通荞麦耐铝性鉴定的适宜条件为发芽种子于500μmol/LAlCl3溶液(pH4.5)处理3d,以发芽种子在这三天内的根伸长量衡量耐铝性程度。在该处理条件下,普通荞麦不同品种间的耐铝性有显著差异。其中,陕西大红花甜荞品种、日本大粒荞、织金红花甜荞的耐铝毒胁迫能力最强,值得在荞麦耐铝性育种和耐铝机制研究中利用。     

The biosynthesis of flavonoid pigments: On the incorporation of phloroglucinol and phloroglucinyl cinnamate into rutin in Fagopyrum esculentum

The problem of whether phloroglucinol is a direct biosynthetic precursor of flavonoids was reinvestigated. Phloroglucinol-2,4,6-¹⁴C was found to be incorporated into rutin in Buckwheat (Fagopyrum esculentum) but most of the activity was found in the sugar moiety, the remainder being approximately equally distributed among the A- and B-rings of the aglycone, quercetin. This indicates extensive degradation of the added phloroglucinol prior to its utilization in the biosynthesis of the flavonoid. The hypothesis of a bio-Fries rearrangement of phloroglucinyl cinnamate to a chalcone, and hence to flavonoids, was also eliminated by comparing the efficiency of incorporation of ¹⁴C-labelled phloroglucinyl cinnamate and those of labelled phloroglucinol and cinnamic acid.     

Carotene epoxides of Lycopersicon esculentum

A series of oxygenated carotenoids has been isolated from tomatoes. Two of these compounds have been identified, by comparison of their chromatographic and spectroscopic properties with those of semisynthetic samples, as epoxides of lycopene (1,2-epoxy-1,2-dihydro-ψ,ψ-carotene and 5,6-epoxy-5,6-dihydro-ψ,ψ-carotene). The other related compounds have been identified by their chromatographic, spectroscopic and chemical properties as mutatochrome (5,8-epoxy-5,8-dihydro-β,β-carotene) and epoxides of phytoene (1,2-epoxy-1,2,7,8,11,12,7′,8′,11′,12′-decahydro-ψ, ψ-carotene), phytofluene (1,2-epoxy-1,2,7,8, 11,12,7′,8′-octahydro-ψ,ψ-carotene and 1,2-epoxy-1,2,7,8,7′,8′,11′,12′-octahydro-ψ,ψ-carotene) and ξ-carotene (1,2-epoxy-1,2,7,8,7′,8′-hexahydro-ψ,ψ-carotene). The presence in tomatoes of apo-6′-lycopenal (6′-apo-ψ-caroten-6′-al), 8′-apo-lycopenal (8′-apo-ψ-caroten-8′-al) and lycoxanthin (ψ,ψ-caroten-16-ol) has been confirmed by comparison with authentic samples.     

Constituants volatils de Lycopersicon esculentum

The volatile components of the tomato have been studied by combined GC-MS with and without previous separation by preparative GC. 5-Methylfurfuryl ketone, furfuryl alcohol, p-anisaldehyde, p-vinylphenol and geraniol were identified and 2-methyl-2-pentanol, 2-heptanol, 4-heptanol, trans-3-hexene-1-ol and trans-2-hexene-1-ol, sec-butyl butyrate, lactone of 5,6-dihydroxyhexanoic acid and isopropyl anisole were tentatively identified.     

二型花柱植株金荞麦繁殖特征

开花物候及繁殖分配是植物适应环境的重要因素。对金荞麦开花物候、繁殖分配及策略进行了研究。结果如下:金荞麦的花果期为每年的8—11月,9月集中开花,其集中开花模式有助于吸引昆虫传粉,提高繁殖成功率;金荞麦单花开花持续时间为1—2 d,种群花期均为85d。L型花序花期为15—26d,S型花序花期为14—27d,两者没有显著差异;L型单花序开花数为26—131朵,S型单花序开花数为36—147朵,两者没有显著差异。L型和S型花序开花动态呈现单峰曲线,在花序开花后第11天L型和S型都达到最大值,分别为7.30%和7.20%,且两种花型具有较高的开花同步性,这有助于其繁殖适应性的提高。同一个花型中,雌蕊长、雄蕊长之间存在极显著负相关,但雌雄总长不存在显著差异,表明雌蕊长、雄蕊长可能存在权衡关系;金荞麦的繁殖器官和营养器官生物量在L型和S型间不存在显著差异,但其花生物量与植株生物量表现出极显著正相关关系。金荞麦L型花生物量分配极显著大于S型,而总生物量不存在显著差异,说明金荞麦植株的营养生长与有性繁殖间存在权衡关系。     

Cloning, Expression, and Mapping of GDP-D-mannose Pyrophosphorylase cDNA from Tomato (Lycopersicon esculentum)

GDP-D-mannose pyrophosphorylase (GMP, EC 2.7.7.22) catalyzes the synthesis of GDP-D-mannose and represents the first committed step in plant ascorbic acid biosynthesis. Using potato GMP cDNA sequence as a querying probe, 65 highly homologous tomato ESTs were obtained from dbEST of GenBank and the putative cDNA sequence of tomato GMP was assembled. The full-length GMP cDNA of tomato was cloned by RACE-PCR with primers designed according to the assembled cDNA sequence. The full-length cDNA sequence contained a complete open reading frame (ORF) of 1 086 bp, which encoded 361 amino acid residues. This gene was designated as LeGMP (GenBank accession No. AY605668). Homology analysis of LeGMP showed a 96% identity with potato GMP and the deduced amino acid showed 99%, 97%, 91% and 89% homology with GMP from potato, tobacco, alfalfa and Arabidopsis thaliana, respectively. Northern blot analysis showed that LeGMP was constitutively expressed in roots, stems, leaves, flowers and fruits of tomato; but the expression levels varied.LeGMP was mapped to 3-D using 75 tomato introgression lines (ILs), each containing a single homozygous RFLP-defined chromosome segment from the green-fruited species Lycopersicon pennellii.     

Chemosystematic evaluation of Aloe section Pictae (Asphodelaceae)

The chemosystematic value of UV-absorbing leaf constituents was considered in previously uncharacterised representatives of Aloe section Pictae, the problematic maculate species complex. Comparative data indicate that the anthrone C-glycoside, 6′-malonylnataloin (7-hydroxychrysaloin 6′-O-malonate) is typical of maculate species in East Africa, but is unconvincing as a synapomorphy for section Pictae. A naphthalene derivative found widely in Aloe, plicataloside, was detected in Aloe greatheadii. Biogeographical trends were observed in the occurrence of the flavonoids isoorientin (luteolin-6-C-glucoside) and isovitexin (apigenin 6-C-glucoside). Isoorientin is a common constituent of tropical and sub-tropical species of Aloe, whereas isovitexin is restricted to a few southern African species. Isoorientin and isovitexin co-occur in the southern African maculate species Aloe parvibracteata, and the disjunct West African maculate species, Aloe macrocarpa. This is the first report of isoorientin and isovitexin in maculate species of Aloe; the presence of flavonoids in section Pictae is of taxonomic interest.     

Bioaccumulation du Cd et du Zn chez les plants de tomates (Lycopersicon esculentum L.)

This work aims at evaluating the accumulation of cadmium (Cd) and zinc (Zn) (trace elements) in the organs of young tomato plants (Lycopersicon esculentum L. var. Rio Grande) and their effects on the rate of chlorophyll and enzyme activities involved in the antioxidant system: catalase (CAT), glutathion-S-transferase (GST) and peroxysase ascorbate (APX). Plants previously grown on a basic nutrient solution were undergoing treatment for 7 days, either by increasing concentrations of CdCl₂ or ZnSO₄ (0, 50, 100, 250, 500 μM) or by the combined concentrations of Cd and Zn (100/50, 100/100, 100/250, 100/500 μM). The results concerning the determination of metals in the various compartments of tomato plants as a function of increasing concentrations of Cd or Zn, suggest a greater accumulation of Cd and Zn in the roots compared to leaves. The combined treatment (Cd/Zn) interferes with the absorption of the two elements according to their concentrations in the culture medium. The presence of Zn at low concentrations (50 μM of Zn/100 μM Cd) has little influence on the accumulation of Cd in the roots and leaves, while the absorption of these two elements in the leaves increases and decreases in roots when their concentrations are equivalent (100/100 μM) compared to treatment alone. When the concentration of Zn is higher than that of Cd (500 μM of Zn/100 μM Cd) absorption of the latter is inhibited in the roots while increasing their translocation to the leaves. Meanwhile, the dosage of chlorophylls shows that they tend to decrease in a dose-dependent for both treatments (Cd or Cd/Zn), however, treatment with low concentrations of Zn (50 and 100 μM) stimulates chlorophyll synthesis. However, treatment with different concentrations of Cd seems to induce the activity of the enzymes studied (CAT, APX, GST). It is the same for treatment with different concentrations of Zn and this particularly for the highest concentrations. Finally, the combined treatment (Zn/Cd) also appears to cause enzyme inductions: CAT, APX and GST.     

Electrophoretic investigation of enzymes from developing Lycopersicon esculentum fruit

Acetone powders were prepared from tomato fruit tissue sampled during development and the proteins were separated by polyacrylamide disc gel electrophoresis. The gels were stained to show up general proteins, lipoproteins, glycoproteins and certain enzymes. Minor changes in protein and glycoprotein patterns accompanied development. Most enzymes exhibited more than one active band, with maximum diversity and specific activities usually appearing in extracts from mature green tissue and least with over-ripe tissue. The results support the view that enzyme synthesis accompanies the climacteric respiration rise at the expense of non-metabolic protein.     

Some properties of pyruvate kinase extracted from Lycopersicon esculentum

Pyruvate kinase was extracted from Me₂CO-dried tissue of various parts of tomato plants. Recovery of the enzyme was improved by the inclusion of thiols in the extraction medium, and its stability was increased considerably in the presence of glycerol and to a lesser extent tetramethylammonium chloride. A phosphatase was present in the tissue extracts which hydrolyses phosphoenolpyruvate in the absence of added ADP. ATP inhibited pyruvate kinase but stimulated the phosphatase, while Mg²⁺ stimulated both enzymes. Data obtained suggest that tomato leaf pyruvate kinase has an absolute dependence on monovalent cations for activity, K⁺ being the principal activator. The phosphatase was inhibited non-selectively by monovalent cations. The total activity of pyruvate kinase and its concentration on a tissue fresh weight basis was greatest in the leaves, activity increasing with the maturity of the tissue. Less enzyme was present in roots, and least in the fruit.     

海蜇养殖对池塘底泥营养盐和大型底栖动物群落结构的影响

2009年5-9月,对荣成靖海湾大型养殖池塘海蜇(Rhopilema esculentum Kishinouye)养殖期间与养殖期前后底泥营养盐及大型底栖动物群落结构变化进行了研究。结果表明,7月份海蜇养殖期间,由于海蜇的避光性而产生的上下浮动的行为特征造成了对水体的扰动作用, 与海蜇的生物沉积作用共同导致养殖海区(实验点)与邻近非养殖海区(对照点)之间各项底泥营养盐指标均存在显著性差异,其中,实验点氨氮(NH₄-N)、硝氮(NO₃-N)和沉降速率(sedimentation rate, SR)显著高于对照点,实验点叶绿素a(Chla)、总有机物(TOM)和总有机碳(TOC)含量显著低于对照点。多变量聚类分析结果表明,海蜇养殖对养殖池塘的大型底栖动物群落结构产生显著影响,并且7月份实验点大型底栖动物生物多样性指数(H')和均匀度指数(J) 随海蜇放养显著增大,并显著高于对照点。大型底栖动物群落多样性指数与沉降速率(SR)和底泥TOM含量分别表现出显著正相关和负相关,而与其他营养盐指标无显著相关性。     

Glutamate oxaloacetate transaminase activity in developing Lycopersicon esculentum fruit

Glutamate oxaloacetate transaminase (GOT) occurs in both the mitochondrial and cytoplasmic fractions from tomato fruit tissue. Changes in activity of the enzyme from fruit at selected developmental stages have been followed. The combined activity fell from the immature green stage to the full red condition whilst the proportion in the mitochondria reached a peak in green-orange fruit. The activity of cytoplasmic, but not mitochondrial, GOT was stimulated by the addition of pyridoxal-5-phosphate. In the green areas of fruit showing blotchy ripening, the combined activity was equivalent to that in normal immature green fruit but with a much higher proportion of the activity in the mitochondria. Mitochondrial GOT could constitute a system in ripening tomato fruit whereby the accumulation of inhibitory concentrations of oxaloacetate affecting the oxidation of succinate and malate might be controlled.     

Sucrose absorption and synthesis by excised Lycopersicon esculentum roots

Asymmetrically-labelled sucrose was absorbed intact by excised roots of tomato, grown in sucrose. Glucose-grown roots possessed sucrose synthetase and sucrose phosphate synthetase activity.     

Isolation of signaling mutants of tomato (Lycopersicon esculentum)

As a first step towards developing a genetic system for investigating signaling processes in plants, we have developed a screen for signaling mutants deficient in a wound response. We have isolated two mutants of tomato that lack detectable production of proteinase inhibitors induced systemically in leaves by wounding. The mutants are deficient in the induction of both proteinase Inhibitor I and proteinase Inhibitor II but can be induced to respond at near wild-type levels by methyl jasmonate, a known elicitor of inhibitor production in tomato. While completely deficient in systemic production of proteinase inhibitors, both mutants produce some proteinase inhibitor in wounded leaves. This evidence suggests the existence of two signaling pathways, one local and one systemic, that regulate the induction of proteinase inhibitor snythesis in response to wounding.     

Purification and properties of phosphofructokinase from fruits of Lycopersicon esculentum

A procedure is described using affinity chromatography on Blue Sepharose and on an immobilized ATP column by which phosphofructokinase has been purified by 260-fold from tomato fruits. The properties of the enzyme are affected by the pH at which the preparation is made and maintained. At the pH optimum, pH 8.0, the enzyme is very heterogeneous with up to three forms present differing in MW. At pH 7.5 a single major form of MW 180 000 is present, and evidence that raising the pH to 8.0 promotes dissociation of the enzyme is discussed.     

Pigment evolution in Lycopersicon esculentum fruits during growth and ripening

During growth and subsequent maturation, the distribution and formation of pigments in the inner pulp and in the outer region of the pericarp of ‘che