甜菜氮素同化与蔗糖代谢的关键酶及相关性研究

试验以甜菜高糖品种甜研7号为试材,采用框栽方法,进行各生育时期氮素同化酶与蔗糖代谢关键酶的测定。研究表明,在甜菜幼苗期和叶丛形成期,叶片硝酸还原酶（NR）活性分别与根中蔗糖磷酸合成酶（SPS）活性和蔗糖合成酶（SS）分解方向活性呈显著和极显著关系,叶片NR活性与根中SS合成方向活性关系虽呈正相关但未达到显著水平;在甜菜块根增长初期,根中谷氨酰胺合成酶（GS）活性与根中SPS活性呈正相关但未达显著水平、分别与根中SS合成与分解方向活性呈显著和极显著关系。可见根中氮素同化和蔗糖代谢存在既相互联系又相互制约的关系。     

甜菜氮素同化与蔗糖代谢的关键酶及相关性研究

试验以甜菜高糖品种甜研7号为试材,采用框栽方法,进行各生育时期氮素同化酶与蔗糖代谢关键酶的测定。研究表明,在甜菜幼苗期和叶丛形成期,叶片硝酸还原酶(NR)活性分别与根中蔗糖磷酸合成酶(SPS)活性和蔗糖合成酶(SS)分解方向活性呈显著和极显著关系,叶片NR活性与根中SS合成方向活性关系虽呈正相关但未达到显著水平;在甜菜块根增长初期,根中谷氨酰胺合成酶(GS)活性与根中SPS活性呈正相关但未达显著水平、分别与根中SS合成与分解方向活性呈显著和极显著关系。可见根中氮素同化和蔗糖代谢存在既相互联系又相互制约的关系。     

氮素用量对寒地水稻氮代谢关键酶活性的影响

本试验以寒地水稻松粳6号和松粳9号为材料,研究了氮素用量对寒地水稻功能叶片氮代谢关键酶活性的影响。研究结果表明:寒地水稻功能叶片NR、GS活性在整个生育期均呈单峰曲线变化,峰值均出现在抽穗期,氮素用量对功能叶片NR、GS活性的影响因品种和时期而异;寒地水稻功能叶片RuBP羧化酶活性从齐穗后呈下降趋势,并且随施氮量的增加而提高;不同生育时期NR、GS、RuBP羧化酶活性与产量及产量构成因子相关关系和相关程度不一致;在一定范围内,水稻产量随着氮量增加而增加,氮素用量与产量间呈一元二次曲线关系。     

节瓜生育过程RubisCO活性与氮钾营养的关系

研究了节瓜生育过程中叶片核酮糖－１,５－二磷酸羧化酶／加氧酶（ＲｕｂｉｓＣＯ）活性的变化及其与氮钾营养的关系氮钾营养设置低氮低钾（Ｎ１２Ｋ９）、低氮中钾（Ｎ１２Ｋ１８）、低氮高钾（Ｎ１２Ｋ３６）、高氮低钾（Ｎ１８Ｋ９）、高氮中钾（Ｎ１８Ｋ１８）和高氮高钾（Ｎ１８Ｋ３６）等６个处理结果表明：在节瓜的生育过程,叶片的ＲｕｂｉｓＣＯ羧化活性和羧化／加氧活性比不断提高,加氧活性则不断降低盛收期植株第１１－２０节的低节位叶片的ＲｕｂｉｓＣＯ羧化活性和加氧活性都最高,羧化／加氧活性比较高;第２１－３０节的中节位叶片的羧化活性较高,加氧活性最低,羧化／加氧活性比最高;第３１－３５节的高节位叶片的羧化活性最低,加氧活性较高,羧化／加氧活性比最低无论氮水平高低,中钾比低钾和高钾都提高ＲｕｂｉｓＣＯ羧化活性和羧化／加氧活性比在３种钾水平下,高氮比低氮降低ＲｕｂｉｓＣＯ羧化活性和羧化／加氧活性比,提高加氧活性     

蛋白核酸合成抑制剂对甜菜谷氨酰胺合成酶活性调控分析

在氮素诱导下,进行甜菜谷氨酰胺合成酶(GS)活性的测定,筛选出最佳的氮素诱导处理,研究核酸合成抑制剂放线菌素D和蛋白合成抑制剂放线菌酮对GS在酶活水平上的表达调控。结果表明,NO3-:NH4+=80:20处理时,GS活性最高,并且在此处理下放线菌素D和放线菌酮均能抑制GS活性。说明甜菜GS的表达可能在转录水平和翻译水平上均受到调控。     

低温诱导甜菜(Beta vulgaris L.)抽薹相关基因的RACE分析

采用cDNA的末端快速扩增(Rapid amplificationn of cDNA ends)技术,以代表性差异分析cDNA-RDA(Representational difference analysis of cDNA)技术获得的低温诱导甜菜茎尖中差异表达的基因片段为模板,进行巢式PCR扩增,然后根据RACE拼接结果合成引物,分别以低温诱导的cDNA和DNA为模板进行PCR扩增,克隆测序后获得1个全长609 bp的cDNA和855 bp的DNA,分别命名为Ty7Br600和Ty7Br900,在GenBank注册,登录号分别为AY324115和AY324114。在GenBank库中的核苷酸序列比较未发现它们的同源序列,因此认为这个cDNA序列可能是甜菜的一个新基因。     

Studies on some characteristics of nitrate reductase from sugar beet （Beta vulgaris L.）leaves

Some characteristics of nitrate reductase from sugar beet leaves shown in this paper were as follows:The nitrate reductase from sugar beet leaves required NADH as an electron donor.Accordingly,the nitrate reductase was classified as NADH-dependent(E.C.1.6.61).The Km value of the nitrate reductase for NADH and NO3^- were 0.86m mol and 0.18μ mol respectively.The optimum pH in reaction mixture solution for nitrate reduction activity was 7.5.The effect of variable concentrations of inorganic phosphorus in the reaction buffer on nitrate reductase activity was investigated.When the inorganic phosphorus concentration was below 35m mol,the nitrate reductase activity was increased with increase of inorganic phosphorus concentration.Conversely,when the inorganic phosphorus concentration was over 35m mol,the nitrate reductase activity was inhibited.The nitrate reductase activity assayed in vitro was 3.2 and 5.6times of that assayed in vivo under the condition of exogenous and endogenous ground substance respectively.     

The Effect of Different Nitrogen Form on Key Enzyme Activity of Sugarbeet（Vulgaris L.） Carbon and Nitrogen Metabolism

This article analyses the effect of the proportion of the different nitrogen forms on key enzyme activity of carbon and nitrogen metabolism under the condition of nutritional water while Tian Yan-7 was used as experimental material.The result showed that nitrate reductase(NR) activity in the leaves gradually enhanced with the increase of NO3^-,No matter in root or leaves,glutamina synthetase(GS) activity first enhanced with increasing NH4^ when NH4^ was lower than that of NO3^-,and GS activity was the highest when NH4^ and NO3^- was equal,then GS activity declined with NH4^ increasing further,In the anaphase of growth,synthetic activity in root of sucrose synthetase(SS) in the mixed NH4^ and NO3^- was obviously highr than or NO3^- alone,Both of the root and sugar yields were the highest when the proportion of NH4^ and NO3^- was 1:1.     

甜菜氮代谢关键酶与其产质量的关系

不同氮肥处理的 NR和 GS与 α-氨基态氮呈线性相关 ,但不同生育期间 ,NR及叶部的 GS与 α-氨基态氮相关不显著 ,而根部的 GS与 α-氨基态氮则呈显著正相关。NR和 GS与产量及产糖量则呈二次型抛物线关系 ,即在一定范围内 ,随 NR和 GS活性增强 ,产量和产糖量增加 ,但超过 NR和 GS的最适值时 ,二者呈下降趋势。在硝态氮作用下 ,NR与各测试指标相关性较大 ,而 GS与各指标在铵态氮处理下相关性较大 ,说明硝态氮对 NR有促进作用 ,而铵态氮对 GS有较强的促进作用。     

低温诱导甜菜(Beta vulgaris L.)Ty7Br600基因的Northern blotting和Southern blotting分析

以低温诱导甜菜幼苗为试验材料,以未进行低温诱导的甜菜幼苗为对照,采用α-32P-ATP放射标记按随机引物标记法标记本实验室克隆的Ty7Br600基因,进行Northern杂交。Northern杂交试验结果显示,在低温诱导培养的甜菜幼苗的RNA群体中,出现较强的杂交条带,而在未诱导培养的甜菜幼苗的RNA群体中,则几乎没有阳性杂交条带出现。因此,Ty7Br600这一序列有可能是二年生甜菜幼苗经低温诱导处理之后被诱导表达的与抽薹相关的新基因序列。Southern杂交结果发现,在每一组酶切中至少有2~3条条带,表明Ty7Br600确为甜菜基因组片段,也同时表明该基因在甜菜基因组中以2个拷贝或低拷贝形式存在。     

Regulation of sucrose synthase activity and sugar yield by nitrogen in sugar beet

The content of sugar is influenced by sucrose synthase (SS) activity in roots. The effects of nitrogen level in the amino- nitrate ratio on SS activity of leaves and roots, roots yield and sugar content in sugar beet were studied in the field experiment by nutrient solution culture. The results showed that SS activity in leaves was lower than that in roots. With nitrogen level increasing, SS decomposition activity enhanced, and synthesis activity reduced. SS activity was regulated by different nitrogen forms and the ratio of NO3– and NH4 . SS synthesis activity was enhanced as NH4 increasing when NO3–: NH4 ≥1, and it decreased as increasing NH4 when NO3–: NH4 ≤1, and it was the highest when NO3–: NH4 =1. SS decomposition activity was enhanced as NO3– increasing. Sucrose content in root was lowed as nitrogen level increasing, but it was enhanced as NH4 increasing in the same nitrogen level. Root and sugar yield were the highest in the medium nitrogen level and NO3–: NH4 =1. The result in field experiment corresponded with that in the nutrient fluid culture. It provides a basis for using reasonably nitrogen fertilizer in sugar beet production.     

高、低糖甘蔗品种叶片转化酶活性与蔗糖代谢关系研究

甘蔗是一种重要的经济作物,在全球蔗糖生产中占主导地位。当前研究者们正致力于通过揭示蔗糖代谢的复杂性而提高蔗茎蔗糖分。为研究探讨甘蔗工艺成熟期不同阶段早熟（高糖）品种和晚熟（低糖）品种的叶片转化酶活性,测定了早熟和晚熟甘蔗品种光合作用很强的第3叶的可溶性酸性转化酶、中性转化酶活性以及叶片酸性转化酶与中性转化酶活性。结果表明,低糖甘蔗品种叶片的酸性转化酶活性较高,而中性转化酶活性较低。相关分析结果表明,叶片酸性转化酶活性与蔗糖分呈负相关,而与己糖含量呈正相关;相反,叶片中性转化酶与蔗糖分呈正相关,与己糖含量呈负相关。研究结果说明,不管甘蔗品种的熟性表现如何,其叶片转化酶在甘蔗糖分调节中均起主导作用。     

嫁接砧木对西瓜果实糖分积累及蔗糖代谢相关酶活性的影响

【目的】研究在果实发育过程中嫁接西瓜糖分积累及蔗糖代谢相关酶活性的变化,探讨不同砧木影响果实品质的机理。【方法】采用优质中小型西瓜早佳为接穗和对照,丝瓜、南瓜(铁木真)、葫芦(昌砧力士F1)、野生西瓜1号、野生西瓜2号为砧木,测定嫁接和对照西瓜果实在发育过程中果糖、葡萄糖、蔗糖的含量以及与蔗糖积累相关酶的活性。【结果】与对照相比,野生西瓜2号/早佳组合的果实糖积累极显著增高,而其他嫁接组合的糖积累均降低,丝瓜/早佳组合的果实糖含量最低;自根和嫁接西瓜果实发育过程中蔗糖转化酶(Inv)活性降低,蔗糖磷酸合酶(SPS)活性升高,蔗糖合酶(SS)在西瓜果实发育后期主要起合成作用,促进果实蔗糖积累,蔗糖代谢相关酶净活性值增大。在整个果实发育过程中,Inv、SPS、SS共同作用于嫁接西瓜和自根西瓜果实的糖分积累及构成;嫁接和对照西瓜果实以己糖积累为主,蔗糖积累为辅。【结论】砧木不同,西瓜果实的糖分积累量也不同,其中野生西瓜2号对早佳西瓜品质影响最小,可以作为优良的嫁接砧木在生产实践中应用。