不同酸度酸雨对小麦花后氮硫代谢和籽粒蛋白组分的影响

酸雨是中国重要的环境问题,为研究酸雨对小麦籽粒品质的可能影响,以小麦品种扬麦15和汶农17为材料开展盆栽试验,研究了不同酸度(pH2.5、p H4.0和p H5.6)酸雨对小麦花后氮硫代谢关键酶活性和籽粒蛋白质含量及组分的影响。结果显示:酸雨处理抑制叶片硝酸还原酶(NR)活性,提高了扬麦15整个灌浆期及汶农17灌浆中后期叶片谷氨酰胺合成酶(GS)活性,促进了叶片蛋白的降解,降低了叶片可溶性蛋白含量。不同酸度酸雨提高了成熟期籽粒中蛋白质含量,酸度越强,增加幅度越大,籽粒中各蛋白组分含量和大部分氨基酸含量也有明显提高。酸雨提高了扬麦15叶片丝氨酸乙酰转移酶(SAT)和O-乙酰丝氨酸硫裂解酶(OAS-TL)活性,但对汶农17硫代谢关键酶活性影响较小,酸雨处理还提高了籽粒中二硫键和含硫氨基酸含量。可见酸雨对小麦氮硫代谢有不同程度影响,进而影响了小麦籽粒蛋白质含量和组成,酸度越强影响越大,但不同品种对酸雨响应有一定差异。     

模拟酸雨对小麦产量及籽粒蛋白质和淀粉含量及组分的影响

以宁麦13和徐麦31两种小麦(Triticum aestivum)品种为材料,通过盆栽试验研究了不同pH值酸雨对小麦产量和籽粒品质的影响。结果表明:模拟酸雨抑制了小麦的生长,减少了生物量的积累。pH值2.0酸雨处理后宁麦13的单穗粒数和单茎产量分别较对照下降了48.6%和56.7%,徐麦31则分别下降了31.2%和39.7%,差异显著。小麦籽粒主要营养成分对酸雨胁迫响应不同,酸雨处理提高了籽粒氨基酸、蛋白质含量,pH值2.0酸雨处理后,宁麦13和徐麦31小麦籽粒中氨基酸含量分别比对照高36.6%和30.9%,总蛋白含量分别比对照高20.6%和15.1%,均与对照差异显著。而小麦可溶性糖、淀粉和脂肪含量较对照降低,且总体表现为酸度增强变化幅度增大。不同蛋白组分也对酸雨胁迫反应不同,酸雨处理提高了籽粒中清蛋白和球蛋白含量,而降低了谷蛋白含量和谷/醇。pH值2.0的酸雨处理后,宁麦13和徐麦31的清蛋白含量较对照分别增加了13.1%和23.9%,但与对照差异不显著。酸雨胁迫降低了总淀粉和支链淀粉含量,宁麦13和徐麦31的pH值2.0酸雨处理总淀粉含量分别较对照下降了11.8%和20.2%,与对照差异显著,但对直链淀粉含量影响不明显。可见酸雨不仅影响小麦的产量,而且对品质也有明显影响。酸雨处理尽管提高了籽粒总蛋白含量,但降低了谷蛋白和谷/醇,降低了其加工品质。     

遮光对小麦籽粒淀粉品质和花前贮存非结构碳水化合物转运的影响

以耐荫品种扬麦158和不耐荫品种扬麦11两个小麦品种为材料,研究了拔节至成熟期遮光对小麦籽粒产量、淀粉含量和淀粉糊化特性的影响,并分析了花前营养器官非结构碳水化合物转运及其与籽粒产量、淀粉含量和淀粉糊化特性的关系.结果表明：遮光条件下小麦花前营养器官中可溶性总糖转运量的下降是造成小麦籽粒产量降低的原因之一;小麦支链淀粉在遮光条件下显著降低而直链淀粉变化不明显,导致小麦籽粒总淀粉含量和支／直显著下降;遮光降低了两品种小麦籽粒淀粉峰值粘度;降低了不耐荫品种扬麦11的低谷粘度,提高了其糊化温度,但对耐荫品种扬麦158的低谷粘度和糊化温度无显著影响.     

花后干旱和渍水对不同品质类型小麦籽粒品质形成的影响

 防雨池栽条件下研究了花后干旱和渍水胁迫对两个不同品质类型小麦（Triticum aestivum）品种籽粒产量和品质形成的影响。结果表明,花后渍水和干旱处理明显降低了小麦籽粒产量和蛋白质产量。在整个灌浆期内干旱处理明显提高了籽粒蛋白质和醇溶蛋白含量,而渍水处理降低了籽粒蛋白质及其组分的积累量。籽粒总淀粉和直链淀粉含量以渍水处理最高,而支链淀粉以对照最高。干旱处理提高了籽粒干、湿面筋含量、沉降值和降落值,而渍水处理降低了上述品质指标。试验表明干旱和渍水胁迫对小麦籽粒蛋白质与淀粉的含量和组分及面粉品质等均有不同程度的影响,从而改变了不同品质类型小麦的籽粒品质。     

花后干旱或渍水逆境下氮素对小麦籽粒产量和品质的影响

 池栽试验条件下,设置渍水、干旱和对照3个水分处理,每个水分处理下设置两个施氮水平 ,研究了花后渍水或干旱逆境下氮素营养对两个不同类型小麦（Triticum aestivum） 品种籽粒产量和品质性状的影响。结果表明,与对照相比,花后渍水或干旱处理显著降低了小麦的千粒重、穗粒数和籽粒产量。在适宜水分和干旱条件下,增施氮肥增加了小麦籽粒产量,而在渍水条件下,增施氮肥降低了产量。干旱处理提高了蛋白质含量,干、湿面 筋含量,沉降值和降落值;而渍水处理则降低了小麦籽粒蛋白质含量和干、湿面筋含量。同 一水分处理下,增施氮肥提高了蛋白质含量,谷蛋白/醇溶蛋白比,支链淀粉含量和支/直链淀粉比。在小麦籽粒主要品质性状上存在显著的水氮互作效应,且水分、氮肥及水氮互作效 应对小麦籽粒产量和品质的影响因品种的不同而异。     

花后盐与渍水逆境对小麦籽粒产量及蛋白质和淀粉积累的影响

采用盆栽试验,研究了花后渍水、盐胁迫和盐渍处理对2个小麦品种（扬麦12和淮麦17）籽粒产量及蛋白质和淀粉积累与组分的影响.结果表明：与对照相比,花后渍水、盐胁迫和盐渍处理显著降低了小麦花前贮藏氮素（花前贮藏干物质）转运量和花后同化氮素（花后同化物）输入籽粒量,从而导致小麦籽粒产量、蛋白质和淀粉产量显著降低,其中盐胁迫和盐渍处理表现更为明显.与对照和渍水处理相比,盐胁迫和盐渍处理显著降低了小麦籽粒蛋白质积累量及谷/醇蛋白比,显著提高了蛋白质组分含量;同时降低了小麦籽粒淀粉积累量、淀粉组分含量及直/支链淀粉比;盐胁迫处理对扬麦12的影响较盐渍处理明显,而盐渍处理对淮麦17的影响较盐胁迫处理明显.渍水条件下小麦籽粒蛋白质和淀粉积累量均下降,除淮麦17谷蛋白和清蛋白含量有所提高外,淮麦17其他蛋白组分含量和扬麦12各蛋白组分含量均下降.     

The altered pattern of amylose accumulation in the endosperm of low-amylose barley cultivars is attributable to a single mutant allele of granule-bound starch synthase I with a deletion in the 5'-non-coding region

Reasons for the variable amylose content of endosperm starch from waxy cultivars of barley (Hordeum vulgare) were investigated. The mature grains of most such cultivars contain some amylose, although amounts are much lower than in wild-type cultivars. In these low-amylose cultivars, amylose synthesis starts relatively late in grain development. Starch granules in the outer cell layers of the endosperm contain more amylose than those in the center. This distribution corresponds to that of granule-bound starch synthase I (GBSSI), which is more severely reduced in amount in the center of the endosperm than in the outer cell layers, relative to wild-type cultivars. A second GBSSI in the barley plant, GBSSIb, is not detectable in the endosperm and cannot account for amylose synthesis in the low-amylose cultivars. The change in the expression of GBSSI in the endosperm of the low-amylose cultivars appears to be due to a 413-bp deletion of part of the promoter and 5'-untranslated region of the gene. Although these cultivars are of diverse geographical origin, all carry this same deletion, suggesting that the low-amylose cultivars have a common waxy ancestor. Records suggest a probable source in China, first recorded in the 16th century. Two further families of waxy cultivars have no detectable amylose in the endosperm starch. These amylose-free cultivars were selected in the 20th century from chemically mutagenized populations of wild-type barley. In both cases, 1-bp alterations in the GBSSI gene completely eliminate GBSSI activity.     

Effects of the activities of key enzymes involved in starch biosynthesis on the fine structure of amylopectin in developing rice (Oryza sativa L.) endosperms

The dynamic changes of the activities of enzymes involving in starch biosynthesis, including ADP-glucose pyrophosphorylase (AGPase), soluble starch synthases (SSS), starch branching enzyme (SBE) and starch debranching enzymes (DBE) were studied, and changes of fine structure of amy- lopectin were characterized by isoamylase treatment during rice grain development, using trans anti-waxy gene rice plants. The relationships between the activities of those key enzymes were also analyzed. The amylose synthesis was significantly inhibited in transgenic Wanjing 9522, but the total starch content and final grain weight were less affected as compared with those of non-transgenic Wanjing 9522 rice cultivar. Analyses on the changes of activities of enzymes involving in starch bio- synthesis showed that different enzyme activities were expressed differently during rice endosperm development. Soluble starch synthase is relatively highly expressed in earlier stage of endosperm de- velopment, whilst maximal expression of granule-bound starch synthase (GBSS) occurred in mid-stage of endosperm development. No obvious differences in changes of the activities of AGPase and SBE between two rice cultivars investigated, except the DBEs. Distribution patterns of branches of amy- lopectin changed continually during the development of rice grains and varied between two rice culti- vars. It was suggested that amylopectin synthesis be prior to the synthesis of amylose and different enzymes have different roles in controlling syntheses of branches of amylopectin.     

玉米灌浆期水分差异供应对籽粒淀粉积累及其酶活性的影响

利用普通玉米(Zay mays)‘掖单22’和高油玉米‘高油115’,研究了灌浆期水分差异供应对籽粒淀粉及其组分积累、相关酶活性动态变化的影响。结果表明,两种类型玉米淀粉积累和酶活性动态变化趋势基本一致,但对水分的反应有差异。缺水提高了‘掖单22’籽粒中淀粉、支链淀粉含量,而直链淀粉含量下降,‘高油115’则是籽粒中的淀粉含量、支链淀粉和直链淀粉含量提高;充分供水使淀粉及其组分产量提高;叶片中蔗糖合成酶(SS)、磷酸蔗糖合成酶(SPS)活性随水分供应水平而提高,尤其在授粉后10~30 d增幅更加明显。充分供水明显提高籽粒中腺苷二磷酸葡萄糖焦磷酸化酶(ADPG-PPase)、尿苷二磷酸葡萄糖焦磷酸化酶(UDPG-PPase)、可溶性淀粉合成酶(SSS)和淀粉粒结合淀粉合成酶(GBSS)活性,缺水使籽粒中酶活性下降较早且迅速;SPS、ADPG-PPase、SSS酶活性对缺水反应比较敏感。     

Physicochemical properties of endosperm and pericarp starches during maize development

Endosperm starch and pericarp starch were isolated from maize (B73) kernels at different developmental stages. Starch granules, with small size (2–4 μm diameter), were first observed in the endosperm on 5 days after pollination (DAP). The size of endosperm-starch granules remained similar until 12DAP, but the number increased extensively. A substantial increase in granule size was observed from 14DAP (diameter 4–7 μm) to 30DAP (diameter10–23 μm). The size of starch granules on 30DAP is similar to that of the mature and dried endosperm-starch granules harvested on 45DAP. The starch content of the endosperm was little before 12DAP (less than 2%) and increased rapidly from 10.7% on 14DAP to 88.9% on 30DAP. The amylose content of the endosperm starch increased from 9.2% on 14DAP to 24.2% on 30DAP and 24.4% on 45DAP (mature and dried). The average amylopectin branch chain-length of the endosperm amylopectin increased from DP23.6 on 10DAP to DP26.9 on14DAP and then decreased to DP25.4 on 30DAP and DP24.9 on 45DAP. The onset gelatinization temperature of the endosperm starch increased from 61.3 °C on 8DAP to 69.0 °C on 14DAP and then decreased to 62.8 °C on 45DAP. The results indicated that the structure of endosperm starch was not synthesized consistently through the maturation of kernel. The pericarp starch, however, showed similar granule size, starch content, amylose content, amylopectin structure and thermal properties at different developmental stages of the kernel.     

High amylose mutants of rice,Oryza sativa L.

Summary Five mutant lines of rice with increased amylose content in starch granules were identified among floury endosperm mutants. The amylose contents of the mutants ranged from 29.4% to 35.4% and were about twice as high as that of the normal counterpart. Starch properties of the high amylose mutants were analyzed by column chromatography, X-ray diffractometry, photopastegraphy and scanning electron microscopy. The high amylose mutants produced longer unit chains of amylopectin than those of the normal counterpart as well as an increased amount of amylose. A X-ray diffractogram of starch in the mutant was characterized by a type B pattern, while that in the normal counterpart showed a type A pattern which is typical for starches of common cereals. The temperatures at the initiation of gelatinization of the mutants were much higher than that for the normal counterpart. The endosperm cells of the mutant were loosely packed with irregular round-shaped starch granules, whereas those of the normal counterpart were densely packed with polyhedral starch granules. Judging from the results obtained, it was concluded that starch properties of the high amylose mutants of rice were similar to those of the amylose-extender (ae) mutant of maize.     

Developmental Changes in the Amylose Content of Endosperm Starch of Barley (Hordeum vulgare L.) during the Grain Filling Period after Anthesis

Large and small starch granules were isolated and characterized from kernels of non-waxy (Bozu) and waxy (Yatomi mochi) barleys at their developmental stages of 8, 16, 28 and 40 days after flowering. The amylose content of the large and small granules of the non-waxy barley starch, as determined by the blue value and enzyme-chromatography, increased with the increasing age of the endosperm. Large granules of the non-waxy barley at any given developmental stage contained more amylose than small granules at the same stage, as in the case of mature non-waxy barley starches. Large granules of either the non-waxy or waxy barleys at any given developmental stage had a lower fraction III: fraction II ratio, one of the structural characteristics of amylopectin, than did small granules of the same cultivar at the same developmental stage. The amylose content in large granules of the waxy barley appeared to increase with the increasing age of the endosperm. The amylose content in small granules of the waxy barley at 8 days after flowering was 10%, although that at 16 and 28 days after flowering and at maturity was only 0~1%.     

Ruminal starch digestion characteristics in vitro of barley cultivars with varying amylose content

Three experiments were conducted to study effects of amylose/amylopectin ratios and starch particle size on ruminal digestion characteristics of barley starch using an automated in vitro gas production system. In Experiment 1, starch digestion characteristics were measured in 12 barley cultivars with different amylose/amylopectin ratios, both as milled grain and as purified starch isolated from the original grain samples. The same 12 cultivars, harvested 1 year later from the same locations, were used in Experiment 2. Gas production was measured in milled samples, and in neutral detergent fibre (NDF) extracted from the same samples. The objective of this approach was to estimate gas production from neutral detergent solubles (NDS) as an approximation of starch. This was done by subtracting the NDF gas curve from the total gas production curve. In Experiment 3, starch digestion characteristics were measured for large and small starch granules from nine of the original cultivars used in Experiment 1. The gas curves obtained were fitted to a three-pool Gompertz model, and the effective rate of digestion (kd) was estimated with a two-compartmental rumen model. In Experiment 1, the effective starch kd for milled barley and purified starch were 0.122 and 0.118/h, respectively. Barley cultivars with low amylose (LA) had a higher effective kd (0.148/h) compared with cultivars with normal amylose (NA) (0.115/h) and high amylose (HA) (0.102/h) (P=0.010). Results obtained with milled barley were supported by the purified barley starch sample results, but differences were smaller and only numerically different. In Experiment 2, the ranking of the amylose groups was consistent with those in Experiment 1 (i.e., LA > NA > HA) (P=0.096). However, these differences were not reflected in the effective kd for the NDS fraction (P=0.366). Thus, factors other than those related to starch per se, or other structural features, are apparently important. Barley cultivars in the LA group had a higher effective kd for aNDF (0.098/h) than did NA and HA barley (0.060 and 0.055/h, respectively). Thus, the effect of the amylose group on the effective kd for aNDF corresponded well with the milled barley results. The NDF fraction, directly or indirectly, has a clear impact on the ruminal digestion rate of barley starch. There was no difference in the effective kd for starch between the small (0.126/h) and large (0.129/h) starch granules.     

GBSS‐BINDING PROTEIN,encoding a CBM48 domain‐containing protein,affects rice quality and yield

The percentage of amylose in the endosperm of rice (Oryza sativa) largely determines grain cooking and eating qualities. Granule‐bound starch synthase I (GBSSI) and GBSSII are responsible for amylose biosynthesis in the endosperm and leaf, respectively. Here, we identified OsGBP, a rice GBSS‐binding protein that interacted with GBSSI and GBSSII in vitro and in vivo. The total starch and amylose contents in osgbp mutants were significantly lower than those of wild type in leaves and grains, resulting in reduced grain weight and quality. The carbohydrate‐binding module 48 (CBM48) domain present in the C‐terminus of OsGBP is crucial for OsGBP binding to starch. In the osgbp mutant, the extent of GBSSI and GBSSII binding to starch in the leaf and endosperm was significantly lower than wild type. Our data suggest that OsGBP plays an important role in leaf and endosperm starch biosynthesis by mediating the binding of GBSS proteins to developing starch granules. This elucidation of the function of OsGBP enhances our understanding of the molecular basis of starch biosynthesis in rice and contributes information that can be potentially used for the genetic improvement of yield and grain quality.     

The assessment of two barley starch mutants as potential parents in a malting quality breeding programme

Grain and malt quality parameters were assessed in the barley starch mutants Waxy Hector, characterised by high levels of amylopectin, and Chalky Glenn, which has lines of cleavage across the large starch granules. Results were compared with those of their parental cultivars and suggested differences in the contribution of endosperm cell walls to milling energy. Although hot water extracts were not improved, other components, especially those associated with the fractured starch mutant, could be used in breeding programmes to improve aspects of quality, if they could be disassociated from deleterious effects of the starch mutation, e.g. poor grain filling.     

The influence of amylose on starch granule structure

Starch granules are principally composed of the two glucose polymers amylose and amylopectin. Native starch granules typically contain around 20% amylose and 80% amylopectin. However, it is possible to breed plants that produce starch with very different amylose and amylopectin contents. At present, the precise structural roles played by these two polymers are incompletely understood. In this study, small-angle X-ray scattering techniques have been applied to investigate the effect of varying amylose content on the internal structure of maize, barley and pea starch species. The results suggest that amylose disrupts the structural order within the amylopectin crystallites.     

Gene expression of the biosynthetic enzymes and biosynthesis of starch during Rice Grain development

Amylose and amylopectin are determinants of the physicochemical properties for starch and grain quality in rice. Their biosynthesis is catalyzed by the interplay of ADP-glucose pyrophosphorylase (AGPase), granule-bound starch synthase (GBSS), soluble starch synthase (SSS), a starch branching enzyme (SBE), and a starch debranching enzyme (SDE). In this study, the genes for these enzymes were highly expressed 7 to 28 days after flowering during grain development, and their expression closely matched increases in both starch content and grain weight Among all the tested cultivars, amylose contents in the rice grains remained essentially constant throughout their development The AGPase gene was highly expressed in the high-yield cultivars of both glutinous and non-glutinous rice. The SSS gene was actively expressed when mature GBSS mRNA decreased. Genes responsible for amylopectin biosynthesis were simultaneously expressed in the late stage of grain development. We have now demonstrated that the expression patterns of starch biosynthetic genes differ between glutinous and non-glutinous rice, and between Tongil (a Japonica/ Indica hybrid) and Japonica types.     

Nature of Lysolecithin in Rice Grains

Problems of the interaction between starch and lysolecithin are treated in this article. The endosperm of rice grains was fractionated and it was found that lysolecithin was associated with starch granules. The association was not a mere adsorption, but rather a more firm combination. It was demonstrated that lysolecithin had an ability to combine with starch in aqueous solution, and to cause amylose to precipitate, but not amylopectin. The lysolecithin-amylose complex was isolated, and it was shown that lysolecithin is most certainly included in the helicoidal structure of amylose chain. A possible role of lysolecithin during the course of the starch formation in rice grains is discussed.     

New Starch Phenotypes Produced by TILLING in Barley

Barley grain starch is formed by amylose and amylopectin in a 1∶3 ratio, and is packed into granules of different dimensions. The distribution of granule dimension is bimodal, with a majority of small spherical B-granules and a smaller amount of large discoidal A-granules containing the majority of the starch. Starch granules are semi-crystalline structures with characteristic X-ray diffraction patterns. Distinct features of starch granules are controlled by different enzymes and are relevant for nutritional value or industrial applications. Here, the Targeting-Induced Local Lesions IN Genomes (TILLING) approach was applied on the barley TILLMore TILLING population to identify 29 new alleles in five genes related to starch metabolism known to be expressed in the endosperm during grain filling: BMY1 (Beta-amylase 1), GBSSI (Granule Bound Starch Synthase I), LDA1 (Limit Dextrinase 1), SSI (Starch Synthase I), SSIIa (Starch Synthase IIa). Reserve starch of nine M3 mutant lines carrying missense or nonsense mutations was analysed for granule size, crystallinity and amylose/amylopectin content. Seven mutant lines presented starches with different features in respect to the wild-type: (i) a mutant line with a missense mutation in GBSSI showed a 4-fold reduced amylose/amylopectin ratio; (ii) a missense mutations in SSI resulted in 2-fold increase in A:B granule ratio; (iii) a nonsense mutation in SSIIa was associated with shrunken seeds with a 2-fold increased amylose/amylopectin ratio and different type of crystal packing in the granule; (iv) the remaining four missense mutations suggested a role of LDA1 in granule initiation, and of SSIIa in determining the size of A-granules. We demonstrate the feasibility of the TILLING approach to identify new alleles in genes related to starch metabolism in barley. Based on their novel physicochemical properties, some of the identified new mutations may have nutritional and/or industrial applications.     

The molecular deposition of transgenically modified starch in the starch granule as imaged by functional microscopy

The molecular deposition of starch extracted from normal plants and transgenically modified potato lines was investigated using a combination of light microscopy, environmental scanning electron microscopy (ESEM) and confocal laser scanning microscopy (CLSM). ESEM permitted the detailed (10 nm) topographical analysis of starch granules in their hydrated state. CLSM could reveal internal molar deposition patterns of starch molecules. This was achieved by equimolar labelling of each starch molecule using the aminofluorophore 8-amino-1,3,6-pyrenetrisulfonic acid (APTS). Starch extracted from tubers with low amylose contents (suppressed granule bound starch synthase, GBSS) showed very little APTS fluorescence and starch granules with low molecular weight amylopectin and/or high amylose contents showed high fluorescence. Growth ring structures were sharper in granules with normal or high amylose contents. High amylose granules showed a relatively even distribution in fluorescence while normal and low amylose granules had an intense fluorescence in the hilum indicating a high concentration of amylose in the centre of the granule. Antisense of the starch phosphorylating enzyme (GWD) resulted in low molecular weight amylopectin and small fissures in the granules. Starch granules with suppressed starch branching enzyme (SBE) had severe cracks and rough surfaces. Relationships between starch molecular structure, nano-scale crystalline arrangements and topographical-morphological features were estimated and discussed.