小麦旗叶Rubisco周转与籽粒含氮量的关系

随着旗叶的衰老,Rubisco含量逐渐减少,延缓小麦旗叶的衰老进程（抽穗期施氮肥）,可增加旗叶Rubsco的含量,提高籽粒的全氮含量,在小麦旗叶全展后28天内,Rubisco的^15N丰度处于较高水平,表明仍有Rubisco的重新合成,而在28天以后,Rubisco的15N丰度处于低水平,表明无Rubisco的重新合成,但这时籽粒的15N丰度却上升,旗叶全展后14天的Rubisor 15N丰度高于旗叶中全氮的丰度,说明此时期Rubisco合成的速率高于其它蛋白质,旗叶衰老过程中Rubisco的15N丰度的净转移高于全氮,Rubisco净N转移也高于全氮,表明Rubisco向籽粒中转移的氮素多于其它蛋白质,对籽粒含氮量的影响最大。     

氮、钾水平对小麦籽粒蛋白质合成关键酶活性的影响

在大田栽培条件下,以蛋白质含量差异的两个冬小麦(Triticum aestivum L.)品种宁麦9号(低蛋白)和扬麦10号(中蛋白)为材料,研究了不同氮、钾施肥水平对小麦旗叶与籽粒中游离氨基酸含量、蛋白质合成关键酶活性和籽粒蛋白质含量的影响及其与开花期旗叶氮、钾营养的关系.结果表明,氮、钾施肥显著提高了小麦花后旗叶与籽粒中游离氨基酸和籽粒蛋白质含量、旗叶谷氨酰胺合成酶(GS)及籽粒GS和谷丙转氨酶(GPT)活性,其中氮肥的作用大于钾肥、氮钾肥配合达到最大,对扬麦10号的效应大于宁麦9号.开花期旗叶氮/钾比随施氮水平的增加呈二次曲线变化,随施钾水平的增加呈下降趋势.旗叶GS活性和成熟期籽粒蛋白质含量与氮/钾比均呈显著或极显著的二次曲线关系.因此,旗叶和籽粒中蛋白质合成关键酶活性受开花期旗叶氮/钾比的显著影响,是氮、钾素影响小麦籽粒蛋白质形成的重要生理原因.     

不同形态氮素对专用型小麦花后氮代谢关键酶活性及籽粒蛋白质含量的影响

采用盆栽方法研究了氮素形态对不同专用型小麦开花后氮素同化关键酶活性及籽粒蛋白质含量的影响。结果表明:不同专用型小麦氮素同化关键酶硝酸还原酶、谷氨酰胺合成酶和谷氨酸合酶对氮素形态的反应不同。强筋小麦豫麦34施用酰胺态氮对旗叶硝酸还原酶和谷氨酰胺合成酶活性、籽粒谷氨酰胺合成酶和谷氨酸合酶活性具有明显的促进作用,最终籽粒蛋白质含量较高;中筋小麦豫麦4 9在施用铵态氮时,3种氮素同化关键酶活性均有较大增强,籽粒蛋白质含量最高;弱筋小麦豫麦5 0硝酸还原酶活性以铵态氮处理最高,而籽粒和旗叶谷氨酰胺合成酶和谷氨酸合酶活性在酰胺态氮处理下明显增强,酰胺态氮对籽粒中蛋白质含量的增加具有明显的促进作用。相关性分析表明,籽粒蛋白质含量与旗叶GS活性和籽粒GOGAT活性呈显著或极显著正相关,与旗叶NR活性和GS活性、籽粒GOGAT活性相关性不显著     

氮素形态对小麦花后不同器官内源激素含量的影响

采用盆栽方法,研究了酰胺态氮、铵态氮和硝态氮对小麦(Triticum aestivum)花后根系、旗叶和籽粒内源激素IAA、GA₃、ABA和ZR含量的影响。结果表明,小麦不同器官的内源激素含量对3种氮素形态的响应不同。氮素形态调节籽粒灌浆是通过根系、旗叶和籽粒中内源激素的协同作用而实现的。酰胺态氮与硝态氮处理相比,小麦花后5~15 d,旗叶GA₃含量、籽粒IAA和ABA含量较高,籽粒灌浆速率(Grain-filling rate, GFR)较高;花后15~25 d,根系GA₃含量、旗叶IAA和GA₃含量、籽粒ABA含量较高,籽粒IAA含量较低,GFR较低。铵态氮与硝态氮处理相比,小麦花后5 d,籽粒ZR含量较高;花后15 d前后,籽粒IAA、ABA含量较低,GFR较低;花后20~25 d,根系ZR、GA₃含量较低,旗叶IAA、GA₃含量较低,ABA含量较高,籽粒ABA、GA₃含量较低,IAA含量较高,GFR较高。铵态氮比硝态氮处理的小麦籽粒粒重显著增加。铵态氮和酰胺态氮处理比硝态氮处理增产显著。建议在‘豫麦49’施肥时,使用铵态氮或酰胺态氮并配施硝化抑制剂。     

烯效唑干拌种对小麦氮素积累和运转及籽粒蛋白质品质的影响

通过田间试验,研究了不同烯效唑干拌种剂量对3个不同筋力小麦品种植株氮素积累、运转和籽粒蛋白质品质的影响,结果表明,基因型、环境及烯效唑处理对小麦品质的影响效应依次减小,且均达到了极显著水平,但三者的互作效应较小。烯效唑处理后提高了不同生态点下不同小麦品种籽粒蛋白质含量和产量,处理后的面筋含量和沉淀值增加,面团形成时间和稳定时间延长;干拌种增加了开花期各营养器官中的氮素含量和单株氮素积累量,花后氮素总转移量、总转移率及其对籽粒氮的贡献率极显著提高,且处理后旗叶中可溶性蛋白质含量在花后15 d内均显著高于对照;对籽粒中氮含量而言,烯效唑处理后提高了灌浆初期籽粒中的非蛋白氮含量,花后5—20 d内均高于对照,灌浆期间籽粒蛋白氮含量均高于对照,因而处理后的粗蛋白质含量变化动态特点为谷底高、回升快。研究认为,烯效唑处理如同基因、环境一样独立影响小麦籽粒品质,而烯效唑处理后提高了开花初期旗叶中的可溶性蛋白质含量和花前营养器官中氮素含量及花后氮素转运量,可能是其提高籽粒非蛋白氮含量、促进籽粒蛋白质含量增加和蛋白质质量提高的重要原因之一,烯效唑干拌种对小麦籽粒蛋白质品质的改善具有广适性。     

氮、磷、钾对豫麦50旗叶蔗糖和籽粒淀粉积累的影响

以豫麦50为对象,探讨了氮、磷、钾对小麦旗叶中蔗糖的积累及相关酶活性以及籽粒中淀粉含量和组分的影响.结果表明,施氮可以增加灌浆前期旗叶中的糖含量,施钾则提高了灌浆后期旗叶中的糖含量,而施磷则对旗叶中的糖含量影响不大.施氮、磷、钾均能增加蔗糖合成酶活性,但它们的作用时间不同：施氮活性增加在籽粒灌浆中期,施磷在灌浆前期,而施钾在灌浆前、中期.但氮亦可增加花后24 d的磷酸蔗糖合成酶活性,施磷增加了灌浆前、中期磷酸蔗糖合成酶活性,施钾则增加了灌浆后期旗叶磷酸蔗糖酶活性.施氮、磷、钾都提高了籽粒中总糖含量,增加籽粒中淀粉含量,其中施钾效果最为明显.施磷提高籽粒中直链淀粉的积累,而施钾则显著提高了籽粒中支链淀粉的含量.     

休闲期深翻覆盖对旱地小麦土壤水分及籽粒蛋白质积累的影响

采用大田试验研究旱地小麦休闲期不同时间深翻不同材料覆盖对土壤水分、籽粒蛋白质形成的影响及其生理机制。结果表明:前茬小麦收获后45 d深翻覆盖较15 d可提高播前120～300 cm土壤蓄水量,而降低开花期0～300 cm(除80 cm土层外)土壤蓄水量;显著提高籽粒蛋白质及其组分含量、蛋白质产量、谷/醇比,提高旗叶GS活性及花后10～15 d、25 d旗叶GOGAT活性,均以渗水地膜覆盖效果较好。结果还表明,播前80～200 cm、开花期260～280 cm土壤蓄水量与旗叶GS、旗叶GOGAT活性显著相关,且与旗叶GS活性关系更密切。籽粒蛋白质组分含量、谷/醇比、蛋白质产量与旗叶GS、旗叶GOGAT活性显著相关,且与旗叶GS活性相关性较密切,尤其是谷/醇比。播前和开花期120～300 cm土壤蓄水量与籽粒蛋白质及其组分含量显著相关,播前60～220 cm、开花期240～300 cm土壤蓄水量与谷/醇比显著相关。总之,休闲期等雨后深翻覆盖有利于蓄积休闲期降雨,有利于提高播前深层土壤水分,最终提高籽粒蛋白质品质及质量,且采用渗水地膜覆盖效果最好。     

施氮量和花后土壤含水量对小麦旗叶衰老及粒重的影响

在防雨池栽培条件下,研究了施氮量和花后土壤含水量对小麦旗叶衰老和粒重的影响.结果表明：各氮肥处理下,小麦旗叶SPAD值、可溶性蛋白质含量、超氧化物歧化酶（SOD）活性、过氧化氢酶（CAT）活性和光合速率（P_n）均表现为：花后土壤含水量60%～70%处理>80%～90%处理>40%～50%处理,小麦旗叶丙二醛（MDA）含量表现为：花后土壤含水量40%～50%处理>80%～90%处理>60%～70%处理,表明花后土壤含水量过高或过低均可导致小麦旗叶早衰,影响籽粒灌浆,降低粒重.在花后相同土壤含水量条件下,旗叶SPAD值、可溶性蛋白质含量、SOD活性、CAT活性和P_n均随施氮量的增加而升高,MDA含量随施氮量的增加而降低,表明增施氮肥可以延缓小麦旗叶衰老,但过量施用氮肥则不利于小麦粒重的提高,尤其是在花后土壤缺水的情况下,施用过多氮肥可导致小麦粒重下降.在小麦生产中可以将施用氮肥和控制花后土壤水分含量相结合,延缓小麦植株衰老,提高粒重.     

灌水时期和灌水量对小麦氮代谢相关酶活性和籽粒蛋白质品质的影响

以高产强筋小麦品种'济麦20'为试验材料,在防雨池栽条件下研究了灌水时期和灌水量对小麦氮代谢相关酶活性和籽粒蛋白质品质的影响.结果表明,随灌水时期增多和总灌水量增加,小麦开花后旗叶硝酸还原酶活性和谷氨酰胺合成酶活性显著增高,旗叶蛋白质水解酶内肽酶(EP)、氨肽酶(AP)、羧肽酶(CP)活性降低;小麦籽粒清蛋白和球蛋白含量增加,而籽粒谷蛋白和醇溶蛋白含量、成熟期籽粒谷醇/清球比值(谷蛋白+醇溶蛋白含量/清蛋白+球蛋白含量)降低;面粉沉降值和面团稳定时间显著降低,籽粒产量显著增加.研究发现,在本试验条件下,小麦全生育期灌水3次处理(底墒水+拔节水+开花水)在灌浆前中期旗叶NR和GS活性、灌浆中后期旗叶EP、CP、AP 活性均较高,且成熟期籽粒蛋白质含量、谷醇/清球比值、面团稳定时间、籽粒产量亦较高,是获得小麦高产优质的最佳灌水处理.     

不同环境条件下小麦氮代谢关键酶活性及籽粒品质

研究了两种环境条件下3个不同蛋白含量小麦品种的氮代谢关键酶活性及籽粒品质的差异.结果表明,龙口试验点的小麦旗叶硝酸还原酶(NR)、谷胺酰氨合成酶(GS)和籽粒谷胺酰氨合成酶活性均显著高于泰安试验点,3个品种间的酶活性顺序均为：济麦20＞优麦3号＞PH971942.优质强筋小麦品种的籽粒综合品质性状在龙口试验点的表现优于泰安试验点. 灌浆期环境因素与小麦籽粒品质和酶活性存在显著的相关性,灌浆期间的较高气温、适当干旱和寡照环境有利于提高小麦籽粒品质.龙口试验点的中、强筋小麦品种和泰安试验点的中筋小麦品种蛋白质含量与旗叶NR和GS活性均达显著正相关.小麦品种用途不同,对环境条件的要求不同,适宜的环境条件提高了氮代谢关键酶的活性,利于改善小麦品质.     

Influence of nitrogen supply and sink strength on changes in leaf nitrogen compounds during senescence in two wheat cultivars

Changes in various nitrogen compounds during senescence of the fourth leaf were studied in two cultivars of spring wheat (Triticum aestivum L.). One of the cultivars (Yecora) was supplied with two N levels; the other (Tauro) was grown with the high N level and pruned above the fourth leaf, whereas the control was left intact. In both cultivars grown with high N supply, net nitrogen export from the fourth leaf did not occur until 35 days after sowing (DAS). Loss of leaf soluble proteins started earlier than that of chlorophylis, and coincided initially with an increase in insoluble protein. In N deficient plants the level of total N, soluble protein, and the activity of nitrate reductase (NRA. EC 1.6.6.1) started to decrease about 5 days earlier, and along with chlorophyll, continued to decrease at a faster rate, than in high N plants. Also, with low N supply, the large subunit (LSU, 58 kDa) of ribulose-1.5-bisphosphate carboxylase/oxygenase (Rubisco, EC 4.1.1.39) decreased in greater proportion than other soluble proteins, while with high N supply the decrease in Rubisco LSU was similar to that of other soluble proteins. Nitrogen deficiency caused a greater decrease in soluble proteins than in insoluble proteins, and NRA relative to soluble proteins. The faster senescing Tauro cultivar had lower levels of most parameters, especially NRA, soluble protein and, after 35 DAS. Rubisco LSU as a proportion of soluble protein. The decrease in sink strength due to shoot pruning did generally not affect the level of the various nitrogenous compounds until 35 DAS; thereafter the levels of most parameters, especially soluble protein, Rubisco LSU and, at late stages of senescence, insoluble protein, were higher in pruned than in control shoots. Thus, shoot pruning slows down senescence. The 56- and 78-kDa polypeptides increased, rather than decreased, with leaf age; the level of these two polypeptides showed a negative relationship with Rubisco LSU (r = -0.933 and r = -0.758, respectively).     

小麦开花后旗叶中蔗糖合成与籽粒中蔗糖降解

在小麦开花后,旗叶中蔗糖磷酸合成酶（SPS）活性在开花后14d内一直维持较高水平,蔗糖合成酶（SS）的活性在开花后14-28d较高,蔗糖的含量与SPS活性呈显著正相关,籽粒中蔗糖合成酶（SS）在开花后28d内一直维持较高的活性;与此相对应,籽粒蔗糖的含量在开花后28d内呈明显的下降趋势。而旗叶和籽粒中SS活性均与籽粒淀粉的积累速率呈极显著正相关。     

Effect of source-sink relations and nitrogen nutrition on senescence and N remobilization in the flag leaf of wheat

The relation between the source-sink ratio and nitrogen nutrition on grain yield of wheat ( Triticum aestivum L. cv. Klein Chamaco) was studied in a greenhouse experiment. Plants were grown until anthesis in pots with soil fertilized with 0.16 mmol N per plant twice a week. At anthesis, all leaves but the flag leaf were excised in a group of plants. In another group the treatment consisted in a similar defoliation plus the longitudinal excision of half the ear, while a third group was left untouched as a control. At the same time, the N supply to half of the plants in each group was interrupted, while the other half continued receiving 16 m M N. The defoliated plants showed a longer functional life of the flag leaf than the control, retaining the chlorophyll, soluble proteins and total reduced nitrogen for a longer time. The ear-excised plants showed an intermediate behavior. The plants with the interrupted N supply showed a faster leaf senescence than the N supplied ones, and this coincided with an increase in the proteolytic activity and nitrogen transport to the ear. However there were no differences in ear weight between the two nitrogen treatments. It is concluded that leaves and ear compete for the nitrogen, and that a low level of carbohydrates in the flag leaf, due to a low source-sink ratio, delays leaf senescence.     

Wheat ear carbon assimilation and nitrogen remobilization contribute significantly to grain yield

The role of wheat ears as a source of nitrogen (N) and carbon (C) in the grain filling process has barely been studied. To resolve this question, five wheat genotypes were labeled with ¹⁵N‐enriched nutrient solution. N remobilization and absorption were estimated via the nitrogen isotope composition of total organic matter and Rubisco. Gas exchange analyses showed that ear photosynthesis contributed substantially to grain filling in spite of the great loss of C due to respiration. Of the total kernel N, 64.7% was derived from the N acquired between sowing and anthesis, while the remaining 35.3% was derived from the N acquired between anthesis and maturity. In addition, 1.87 times more N was remobilized to the developing kernel from the ear than from the flag leaf. The higher yielding genotypes showed an increased N remobilization to the kernel compared to the lower yielding genotypes. In addition, the higher yielding genotypes remobilized more N from the ears to the kernel than the lower yielding genotypes, while the lower yielding genotypes remobilized more N from the flag leaf to the kernel. Therefore, the ears contribute significantly toward fulfilling C and N demands during grain filling.     

Subtilisin-like serine proteases involved in N remobilization during grain filling in wheat

The induction of two subtilisin-like proteases (P1 and P2) associated with stress-induced senescence in young plants was investigated in adult wheat plants during the grain-filling period. Western blot analysis of flag leaf extracts showed that P1 was induced very late in the life cycle of the plants (9 days post-anthesis) and that 7 days later it reached a 2.5-fold increase with respect to the initial value at anthesis. On the other hand, the P2 signal was already detected previous to anthesis and increased soon after anthesis, reaching a fourfold increase by the end of the grain-filling period. The induction of P1 and P2 temporally correlates with the degradation of the Rubisco small and large subunits in the flag leaf, as well as with nitrogen (N) accumulation in the ears. At the same time, a decrease in the endogenous concentration of the cytokinins isopentenyladenine and isopentenyladenosine (iP + iPA) in the leaves was observed. In detached leaves senescing in the dark, the levels of both proteases were affected by 6-benzylaminopurine application: the induction of P1 was completely prevented, whereas the induction of P2 was reduced. Our findings demonstrate that both P1 and P2 are expressed in leaves of adult plants and are induced during natural senescence. These results enable us to postulate their participation in N remobilization to developing grains during monocarpic senescence and their regulation by a cytokinin-mediated mechanism.     

硫氮配施对持绿型小麦氮素运转及叶片衰老的影响

以持绿型小麦品种‘豫麦66号’、‘潍麦8号’及非持绿型品种‘小偃6号’为材料,采用以氮肥为主区硫肥为副区的田间裂区试验,研究了2个施氮水平[纯氮120kg/hm2(N120)、220kg/hm2(N220)]和3个施硫水平[纯硫0kg/hm2(S0)、20kg/hm2(S20)、60kg/hm2(S60)]下植株各部位的含氮量、叶片干鲜重及叶片叶绿素含量的变化,探讨硫氮配施对不同类型小麦氮吸收及衰老的影响。结果表明:在相同处理下,持绿型小麦植株的总含氮量、氮素转运量、叶绿素含量、叶片含水量及穗粒数和千粒重均高于非持绿型小麦。N120处理条件下,不同硫肥处理时持绿型小麦与非持绿型小麦变化趋势相同,开花期茎和叶含氮量及叶绿素含量在S60处理下均低于其他2个硫肥处理,生育后期叶片含水量下降幅度也明显高于其他处理;在N220处理条件下,3个品种开花期叶含氮量、收获期总氮累积量、氮收获指数、叶绿素含量及叶片含水量在S60处理下均高于其他2个处理,其中非持绿型小麦在高硫处理条件下灌浆期的叶绿素含量的增长率明显高于持绿型小麦,而灌浆中后期叶片含水量的下降幅度则明显低于持绿型小麦。研究发现,施用硫肥在氮肥不足时会对小麦植株氮素的吸收利用及叶片衰老等方面产生负面影响,但在氮肥充足时却在氮素的吸收利用、延缓叶片衰老及最终籽粒产量和总生物量等方面表现出正面效应;本实验条件下,220kg/hm2左右施氮量和60kg/hm2左右施硫量有利于各品种小麦生长发育和产量提高;高N和高S水平对于延缓小麦的衰老而言,非持绿性小麦比持绿型小麦更明显。     

水稻抽穗后剑叶衰老过程中光合关键酶的基因表达

协优 930 8、培矮 6 4s/ 9311和汕优 6 3三个超级高产水稻品种抽穗后 ,剑叶中光合速率减小、叶绿素含量下降、SOD活力降低和MDA含量上升的幅度不同 ,显示它们开花结实过程中叶片衰老的速度不同 ,以汕优 6 3衰老最快 ,协优 930 8最慢 ,培矮 6 4s/ 9311居中。Northern杂交和免疫定量研究表明 ,汕优 6 3和培矮6 4s/ 9311剑叶rbcS(Rubisco小亚基基因 )和rca(Ru bisco活化酶基因 )mRNA的表达随抽穗后天数的增加而下降 ,而协优 930 8在抽穗后第 10天才开始下降 ;Rubisco和Rubisco活化酶蛋白含量的下降趋势与rbcS和rcamRNA的变化趋势相似。因此 ,水稻抽穗后剑叶衰老过程中光合能力的下降与光合关键酶基因(rbcS和rca)表达下降密切相关     

Proteome reference maps of vegetative tissues in pea. An investigation of nitrogen mobilization from leaves during seed filling

A proteomic approach was used to analyze protein changes during nitrogen mobilization (N mobilization) from leaves to filling seeds in pea (Pisum sativum). First, proteome reference maps were established for mature leaves and stems. They displayed around 190 Coomassie Blue-stained spots with pIs from 4 to 7. A total of 130 spots were identified by mass spectrometry as corresponding to 80 different proteins implicated in a variety of cellular functions. Although the leaf proteome map contained more abundant spots, corresponding to proteins involved in energy/carbon metabolism, than the stem map, their comparison revealed a highly similar protein profile. Second, the leaf proteome map was used to analyze quantitative variations in leaf proteins during N mobilization. Forty percent of the spots showed significant changes in their relative abundance in the total protein extract. The results confirmed the importance of Rubisco as a source of mobilizable nitrogen, and suggested that in pea leaves the rate of degradation of Rubisco may vary throughout N mobilization. Correlated with the loss of Rubisco was an increase in relative abundance of chloroplastic protease regulatory subunits. Concomitantly, the relative abundance of some proteins related to the photosynthetic apparatus (Rubisco activase, Rubisco-binding proteins) and of several chaperones increased. A role for these proteins in the maintenance of a Rubisco activation state and in the PSII repair during the intense proteolytic activity within the chloroplasts was proposed. Finally, two 14-3-3-like proteins, with a potential regulatory role, displayed differential expression patterns during the massive remobilization of nitrogen.