玉米高赖氨酸含量基因型离体筛选方法与应用

设不同浓度赖氨酸(L)加苏氨酸(T)的培养基, 将opaqure-2等32个玉米自交系种子的胚接种在培养基上发芽, 同时测定种子中游离氨基酸和总氨基酸含量, 检测胚生长对L+T的抗性与种子中赖氨酸含量关系, 筛选鉴定玉米种子中高赖氨酸含量的有效方法。结果表明, opaqure-2对L+T有较强抗性, 子粒中有较高含量游离赖氨酸;普通玉米自交系抗L+T能力较弱, 种子中游离氨基酸含量较低;在旅大红骨类群中检测到对L+T有较强抗性的基因型, 子粒中有较高含量的赖氨酸和其他氨基酸。玉米中存在典型的天冬族氨基酸生物合成的反馈抑制代谢途径, 试管L+T胁迫方法可用于筛选和鉴定玉米高赖氨酸基因型。     

小麦花药液体漂浮离体培养体系的建立与应用

为了建立高效的小麦花药离体培养体系,对6个小麦品种（系）的5个花药离体培养性状进行了鉴定,并利用6种不同时间的低温预处理对两个小麦品种幼穗进行了处理效果的探讨。结果表明,6个品种（系）的出愈及绿苗分化能力均较强,愈伤组织诱导率、绿苗分化率和绿苗产率分别达到116.48%、5.79%和10.25%以上,初步建立了小麦花药液体漂浮离体培养体系,其培养效果好于固体培养方式。6种不同时间的低温预处理培养结果表明,幼穗经4 ℃低温预处理后,愈伤组织诱导率、绿苗分化率及绿苗产率与对照相比均表现下降的趋势,白苗分化率及白苗产率具有增加的趋势,说明未进行低温预处理的适期幼穗培养效果更好。     

麦类作物花药离体培养特性遗传控制研究进展

基因型依赖及白苗再生等问题限制了花药离体培养技术潜力的充分发挥。为了克服这些问题,提高花药离体培养效率,国内外学者采用传统遗传学、细胞学、生理生化、分子标记、基因克隆、转录组学及蛋白组学等方法,在小麦、大麦及小黑麦花药预处理、胚状体和愈伤诱导、绿苗再生及白苗形成遗传控制方面进行了大量的研究。本文对这些研究进行综述,以期为相关研究提供参考。     

灭酶方法对燕麦淀粉和蛋白质体外消化特性的影响

为了解灭酶方法对燕麦淀粉和蛋白质体外消化特性的影响,采用炒制、常压蒸制和远红外烘烤三种灭酶方法处理燕麦籽粒,分析了燕麦全粉的营养成分、黏度特性及淀粉和蛋白质体外消化特性的变化。结果表明,炒制灭酶后燕麦全粉粗脂肪含量提高,而β葡聚糖和总淀粉含量降低(P<0.05);常压蒸制灭酶导致燕麦全粉中蛋白质、粗脂肪、总淀粉含量降低(P<0.05);远红外烘烤灭酶对燕麦营养成分影响不大(P>0.05)。灭酶处理后燕麦全粉的糊化温度均降低,常压蒸制和远红外烘烤提高了燕麦全粉的峰值黏度和最终黏度(P<0.05)。灭酶处理使燕麦全粉中抗性淀粉含量显著增加(P<0.05),对慢速消化淀粉的影响不大(P>0.05)。在蛋白质体外消化中,远红外烘烤后燕麦蛋白体外消化率最高,达到70.05%,而常压蒸制灭酶燕麦的蛋白质消化率最低,为62.80%。说明远红外烘烤灭酶处理对燕麦营养组分影响不大,更加有利于消化。     

离体叶段喷雾法建立小麦白粉病菌对三唑酮的敏感基线及其监测技术

为建立小麦白粉病菌对三唑酮抗药性的离体叶段喷雾法监测标准,采用该方法测定了2013年采自河南、河北省部分麦区的101个小麦白粉病菌野生菌株对三唑酮的敏感性。结果表明,小麦白粉病菌群体对三唑酮敏感性的EC50范围为0.239 8~7.558 5μg·mL-1,平均为3.561 2±1.881 3μg·mL-1;其敏感性均呈连续单峰曲线分布,可作为喷雾防治时小麦白粉病菌对三唑酮杀菌剂的敏感性基线。     

Protein Inhibitors ofAlpha-amylase in Mature and Germinating Grain of Rye (Secale cereale)

The activities of endogenous (R-type) and exogenous acting (D-type) protein inhibitors ofalpha-amylase and the activities ofalpha- and total amylase were determined in milling fractions of rye. High D-type amylase inhibitor activities were detected in the embryo (255 IU/g) and in the endosperm fraction (64·9 IU/g), low inhibitor activities were found in the aleurone layer fraction (25·9 IU/g). The highest R-typealpha-amylase inhibitor activity was found in the aleurone layer fraction (32·6 IU/g), and the lowest value in the epidermis containing fraction (5·0 IU/g). The D- and R-typealpha-amylase inhibitor activities varied with growing conditions. D-type amylase inhibitor activities were found to be high in those samples which grew under drought conditions and low in samples cultivated under wet and cool weather. Higher R-typealpha-amylase inhibitor activities were found in rye genotypes cultivated under wet conditions and lower values under dry weather. There were small variations inalpha-amylase inhibitor activities between sprout-stable and sprout-sensitive rye genotypes. The D- and R-typealpha-amylase inhibitor activities of all varieties were stable during 72 h of germination. Similar soil conditions will therefore lead to differentialalpha-amylase inhibitor activities depending on weather conditions during growth.     

禾生指葡孢霉生物学特性及室内药剂筛选

为了给青稞和燕麦鞘腐病的科学防治提供技术支持,对其病原菌-禾生指葡孢霉的生长温度、适宜培养基种类、适宜pH值、存活时间等生物学特性进行了研究,并选取麦类作物上常用的8种杀菌剂进行了室内药剂筛选。结果表明,禾生指葡孢霉在5~30℃下可生长,适宜生长温度为20~25℃;在PSA、PDA、CA、CMA和OMA培养基平板上均可生长、产孢,适宜病菌生长、产孢的培养基为PDA和PSA;在pH值为4~10的PDA平板上均可生长、产孢,适宜生长的pH值为6~10;在5~10℃冷藏条件下,禾生指葡孢霉存活时间超过5a。在试验浓度(推荐剂量)下,8种杀菌剂对禾生指葡孢霉菌丝生长均具有不同程度的抑制效果,其中50%多菌灵可湿性粉剂(500×)、70%代森锰锌可湿性粉剂(500×)、430g·L-1戊唑醇悬浮剂(5 000×,8 000×)和400g·L-1氟硅唑乳油(5 000×,10 000×)的抑菌效果达100%。     

Quality Effects of Rye (Secale cerealeL.) Chromosome Arm 1RL Transferred to Wheat (Triticum aestivumL.)

Quality effects of rye (Secale cerealeL.) chromosome arm 1RL transferred to wheat (Triticum aestivumL.) were characterised by comparison of a group 1R(1B) substitution lines and 1BL.1RS translocation lines. The experimental materials were sister lines derived from the mating Mironovskaya10/NE7060//NE80413. 1R(1B) substitution lines were identified by the presence of ryeomega- andgamma-secalins among 70% ethanol soluble proteins, combined with the presence of high molecular weight (HMW) secalin proteins in total grain protein extracts. Genes on 1RL reduced grain weight, grain hardness, Mixograph time, Mixograph tolerance and SDS sedimentation volumes. 1RL had no effect on flour yield or grain and flour protein concentrations. The HMW secalin proteins encoded by genes on 1RL most probably caused the decline in dough strength seen in 1R(1B) lines relative to that of 1BL.1RS sister lines. Reduced grain hardness might also be related to the presence of HMW secalins, although a role for additional, unidentified genes on 1RL could not be discounted.     

Comparative Studies of HighMrSubunits of Rye and Wheat. I. Isolation and Biochemical Characterisation and Effects on Gluten Extensibility

The structural features of highM_rglutenin subunits of wheat were compared with those of analogous proteins from rye. Subunits of two rye cultivars (Danko and Halo) and of the wheat cultivar Rektor were isolated from defatted flours by extraction with 50% (v/v) aqueous propan-1-ol under reducing conditions at 60°C followed by precipitation using a 60% concentration of propan-1-ol. The yields of dialysed and freeze-dried subunits were 0·33% and 0·32% (w/w of flour), respectively (rye cultivars), and 0·91% (Rektor). SDS–PAGE revealed that the rye cultivars contained at least five subunits with mobilities corresponding to the x-type subunits of wheat. Separation by RP–HPLC indicated that the rye cultivars did not differ in the qualitative composition of subunits, but in their quantitative proportions. The surface hydrophobicities of the rye subunits were significantly lower than those of wheat subunits. The amino acid compositions of single rye subunits were characterised by high contents of Glx, Gly and Pro, and they were closely related to those of wheat subunits, except that the Glx content was generally lower and the Cys content higher. Notable differences between rye and wheat subunits were found in their contributions to gluten strength. Whereas wheat subunits, reoxidised with potassium bromate and mixed with a standard wheat flour, caused a significant increase in gluten strength, reoxidised rye subunits had the opposite effect.     

黑麦GIF基因家族全基因组鉴定及其表达分析

生长调节互作因子(GRF-interacting factor, GIF)是植物体内一类转录共激活因子,在植物生长发育过程中起重要作用。为挖掘和利用黑麦GIF家族基因,利用同源比对方法对黑麦GIF基因家族进行了鉴定,并系统分析了黑麦与其他禾本科植物及模式植物拟南芥GIF基因的系统发育关系、蛋白结构域、基因结构和共线性关系。结果显示,在黑麦基因组中共鉴定到3个GIF基因家族成员,分布在4R和5R染色体上;系统进化分析发现,黑麦GIF基因家族成员与小麦和大麦的亲缘关系更近。蛋白结构域和基因结构分析发现,GIF基因家族成员在黑麦及其他禾本科植物中的蛋白结构域和基因结构相对保守,均含有4个外显子。共线性分析发现,黑麦ScGIF1基因与其他禾本科植物GIF基因间均存在共线性关系,其中与小麦间共线基因数最多。Ka/Ks分析发现,黑麦GIF基因家族成员在进化中受到纯化选择。此外,在黑麦穗部鉴定到一个特异性高表达的GIF基因(ScGIF1),其编码的蛋白含有多个磷酸化位点,推测ScGIF1蛋白可能是通过磷酸化来调控黑麦麦穗的发育和粒型的。     

玉米子粒和果穗离体培养技术及其应用研究进展

本文对玉米籽粒离体培养技术及其在子粒发育生理研究中的应用进行了评述；并提出了果穗离体培养方法．     

山薯试管零余子的诱导

以山薯无菌苗茎段为试验材料,研究BA、NAA、JA、蔗糖浓度、光周期以及不同培养方式对试管零余子诱导形成的影响.结果表明,MS+2.0mg/L BA+0.1 mg,L NAA+80g/L蔗糖+5.0g/L,琼脂+1.5 g/L活性炭,12 h/d的光照时间(光照强度为2 000 lx)是试管零余子诱导较优的培养条件.经过2个月的培养,试管零余子诱导率达90%以上,零余子发生个数为1.44个/株.蔗糖浓度是影响试管零余子发生的最重要因素,高蔗糖浓度(80 g/L)能促进试管零余子的发生.     

Optimisation of the Selective Extraction of (Glucurono)arabinoxylans from Wheat Bran: Use of Barium and Calcium Hydroxide Solution at Elevated Temperatures

Two approaches were investigated in attempts to obtain a high extraction yield of (glucurono) arabinoxylans from water-unextractable cell wall material (WUS) of wheat bran using saturated barium hydroxide containing 0·26 M sodium borohydride. First, the effect of three pretreatments (autoclave treatment, alkaline peroxide and chlorite delignification) of the WUS prior to extraction appeared to have no effect on the extraction yield. Moreover, modifications to the composition and molecular weight distribution of the (glucurono)arabinoxylans occurred when such treatments were used. Second, the effect of an increasing extraction temperature and concentration of alkali was investigated. Increasing the extraction temperature improved the extraction yield of (glucurono)arabinoxylans from 29% at 20°C to 50% at 95°C. Increasing the barium hydroxide concentration with the temperature resulted in no further improvements in extraction yield up to 70°C. Above this temperature the extraction yield decreased. Substitution of barium hydroxide by calcium hydroxide resulted in lower yields and a lower selectivity; the lower solubility of calcium hydroxide may have been responsible for this. Supplementary experiments to investigate the mechanism of the selectivity of the bivalent hydroxide extraction with addition of sodium borohydride indicated a possible role for borate, derived from borohydride.     

Buckwheat Seed Milling Fractions: Description, Macronutrient Composition and Dietary Fibre

The structure of the mature buckwheat achene and groat is discussed in relation to milling fractions and nutritional composition. Whole groats contain 55% starch, 12% protein, 4% lipid, 2% soluble carbohydrates, 7% total dietary fiber (TDF), 2% ash, and 18% other components (organic acids, phenolic compounds, tannins, phosphorylated sugars, nucleotides and nucleic acids, unknown compounds). The composition of the milling fractions reflects the relative abundance of seed tissues. Starch is concentrated in the central endosperm. Protein, oil, soluble carbohydrates and minerals are concentrated in the embryo. Commercial «Fancy» flour, a light-coloured flour, is mostly central endosperm and contains 75% starch, 6% protein, 1% lipid, 1% soluble carbohydrates, 3% TDF, 1% ash, and 13% other components. Although the embryo traverses the central endosperm, during milling parts of the embryo separate with the aleurone and seed coat in the bran fraction. Bran, with little central endosperm, contains 18% starch, 36% protein, 11% lipid, 6% soluble carbohydrates, 15% TDF, 7% ash, and 7% other components. Buckwheat bran also is a rich source of TDF and soluble dietary fibre (SDF), particularly bran with hull fragments (40% TDF of which 25% is SDF), while bran without hull fragments has 16% TDF of which 75% is SDF.     

Apoptogenic Metabolites in Fractions of the Benthic Diatom Cocconeis scutellum parva

Benthic diatoms of the genus Cocconeis contain a specific apoptogenic activity. It triggers a fast destruction of the androgenic gland in the early post-larval life of the marine shrimp Hippolyte inermis, leading to the generation of small females. Previous in vitro investigations demonstrated that crude extracts of these diatoms specifically activate a dose-dependent apoptotic process in human cancer cells (BT20 breast carcinoma) but not in human normal lymphocytes. Here, a bioassay-guided fractionation has been performed to detect the apoptogenic compound(s). Various HPLC separation systems were needed to isolate the active fractions, since the apoptogenic metabolite is highly active, present in low amounts and is masked by abundant but non-active cellular compounds. The activity is due to at least two compounds characterized by different polarities, a hydrophilic and a lipophilic fraction. We purified the lipophilic fraction, which led to the characterization of an active sub-fraction containing a highly lipophilic compound, whose molecular structure has not yet been identified, but is under investigation. The results point to the possible medical uses of the active compound. Once the molecular structure has been identified, the study and modulation of apoptotic processes in various types of cells will be possible.     

利用Solexa测序技术鉴定苋菜试管开花过程中的miRNAs

利用Solexa测序技术对苋菜试管开花过程中的miRNAs进行筛选和鉴定,使用GO数据库和KEGG代谢途径数据库对靶基因进行功能注释和分类。结果显示:共得到sRNA Unique序列5 457 486条,其中与拟南芥基因组完全匹配有27 596条,仅仅占0.51%;miRNA主要分布在18 nt~25 nt之间,其中长度为24 nt序列数量最多;苋菜试管开花过程中miRNA的表达丰度存在明显差异,主要低丰度表达;总共鉴定了235个已知的苋菜miRNA,构成20个家族,不同家族间成员数量存在巨大差异;苋菜保守miRNA的长度主要集中在21 nt和20 nt;另外总共获得14个候选的新miRNA,其中有8个miRNA含单个基因座,另6个miRNA具备多个基因座;11个候选的miRNA预测到了靶基因,总共对应着约46个基因符合靶标特征。这些靶标参与植物生物体的各个发育进程,在苋菜试管开花过程中对生物过程、细胞组分及分子功能起到重要作用。     

转小麦 TaPDI-A、 TaTRXh-A和 TaPP2Ac-D基因拟南芥植株的主要抗旱性研究

核氧还蛋白（nucleoredoxin,NRX）可通过还原目标蛋白的二硫键来调控其生物活性,在植物的生长发育和抗逆境胁迫中发挥着重要作用。蛋白质二硫键异构酶（protein disulfide isomerase,PDI）、h型硫氧还蛋白（h-type thioredoxin,TRXh）和蛋白磷酸酶2A催化亚基（protein phosphatase 2A catalytic subunit,PP2Ac）是小麦核氧还蛋白TaNRX1的互作蛋白。为了明确TaNRX1互作蛋白的抗旱性功能,本研究在拟南芥中过表达了小麦 TaPDI-A、 TaTRXh-A和 TaPP2Ac-D基因,对野生型和转基因拟南芥的表型和抗旱相关生理指标进行了鉴定。结果表明,干旱胁迫处理后,转 TaPDI-A、 TaTRXh-A和 TaPP2Ac-D基因拟南芥的根长、存活率、脯氨酸含量均大于野生型,离体叶片失水率、丙二醛（MAD）含量均小于野生型。二氨基联苯胺（diaminobenzidine,DAB）对H₂O₂组织定位染色结果表明,干旱胁迫处理后,转 TaPDI-A、 TaTRXh-A和 TaPP2Ac-D基因拟南芥的H₂O₂含量均低于野生型。上述结果说明,TaNRX1的互作蛋白基因 TaPDI-A、 TaTRXh-A和 TaPP2Ac-D增强了拟南芥对于干旱胁迫的抵抗能力。本研究可为小麦抗旱育种提供候选基因和理论基础。     

玉米灌浆期3种鳞翅目害虫的空间分布

采用网格式取样200株玉米,整株剖秆调查亚洲玉米螟、桃蛀螟和棉铃虫在玉米上的数量,用地统计学的方法分析和模拟它们在田间的水平分布;采用生态位理论分析3种害虫在玉米植株上的生态位和种间竞争。结果表明,亚洲玉米螟、桃蛀螟和3种鳞翅目害虫整体在玉米田中的水平分布分别适合球形、高斯、球形模型拟合,均属于聚集分布。Kriging插值模拟图显示,亚洲玉米螟和桃蛀螟为核心分布型;在垂直分布上,雌穗上3种害虫数量最多,占总虫量的69.82%。亚洲玉米螟的基础生态位宽度最大,在整株玉米上都可危害;桃蛀螟则主要在玉米中、上部;棉铃虫基础生态位最窄,只在雌穗附近危害。3种害虫在玉米茎秆上种间竞争激烈,异种害虫无法共存;雌穗上种间竞争程度小于茎秆,异种害虫可以共存。     

引发小麦赤霉病和茎基腐病禾谷镰孢菌的生物防治初探

禾谷镰孢菌(Fusarium graminearum)是小麦上的重要病原真菌。通过致病性测定从江苏小麦茎基腐病病害样本分离菌中筛选到了对小麦赤霉病和茎基腐病都有强致病力的F.graminearum菌株GF1117。为了对F.graminearum进行生物防治,利用稀释涂布平板法和平板对峙法从小麦不同生境中分离筛选到35株对GF1117具有明显拮抗效果的细菌菌株,分别在田间和温室进行了小麦赤霉病和茎基腐病的生物防治试验。结果表明,35株拮抗细菌对小麦赤霉病均有一定的防治效果,且在小麦感病品种和中抗品种上对茎基腐病的防效不同;菌株1-8对两种病害的防效都在45%以上。     

Fusarium Species Synthesize Alkaline Proteinases in Infested Barley

Barley (Hordeum vulgare L.) that is infested with Fusarium head blight (FHB, ‘scab’) is unsuitable for malting and brewing because it may contain mycotoxins and has unacceptable malting quality. Fungal proteinases are apparently often involved in plant-microbe interactions, where they degrade storage proteins, but very little is known about the enzymes that the fungi produce in the infected grain. We have shown previously that one plant pathogenic fungus, Fusarium culmorum, produced subtilisin- and trypsin-like enzymes when grown in a cereal protein medium. To establish whether these proteinases were also synthesized in FHB-infested barley in vivo, field-grown barley was infested as the heads emerged. Extracts were prepared from the grain as it developed and matured and their proteolytic activities were measured with N-succinyl-Ala-Ala-Pro-Phe p -nitroanilide and N-benzoyl-Val-Gly-Arg p -nitroanilide. The heavily infested barleys contained both subtilisin- and trypsin-like activities. These enzymes reacted with antibodies prepared against each of the two F. culmorum proteinases, indicating that those produced in the laboratory cultures and in the field-infested barley were the same. The presence of these proteinases correlated with the degradation of specific buffer-soluble proteins in the infested grains. These enzymes readily hydrolyzed barley grain storage proteins (C- and D-hordeins) in vitro. The presence of these Fusarium proteinases in the barley indicates that they probably play an important role in the infestation, but exactly how and when they function is not clear.