人工老化处理对结球甘蓝种子生理生化特性的影响

以结球甘蓝品种‘冬升’种子为材料,研究高温(40℃)高湿(相对湿度100%)人工老化处理过程中种子的萌发特性、种子浸出液相对电导率和丙二醛、可溶性蛋白、可溶性糖含量以及抗氧化酶活性的变化,以揭示种子劣变的机理。结果表明:(1)人工老化处理甘蓝种子的含水量和不正常苗率均随着老化时间的延长逐渐增加,而种子发芽势、发芽率、发芽指数和活力指数的增加均逐渐降低。(2)随着处理天数的延长,老化处理甘蓝种子的浸出液电导率显著增大,浸出液可溶性糖含量逐渐升高,可溶性蛋白含量表现出显著下降趋势,而种子MDA含量呈先升高后逐渐下降的趋势。(3)在结球甘蓝种子老化进程中,其种子中SOD、POD、CAT活性变化的趋势相似,均随老化程度的加深而逐渐降低;而APX活性在老化处理的最初2d显著增加,第6天显著降低。研究发现,在结球甘蓝种子老化进程中,种子活力和萌发率显著降低,其种子浸出液电导率、可溶性蛋白含量、可溶性糖含量、保护酶活性变化与种子老化及劣变程度密切相关,膜脂过氧化作用可能是引起或加剧种子老化劣变的重要原因之一。     

利用热稳定蛋白特异条带鉴别籼粳稻的方法研究

以85份栽培稻为材料,利用SDS-聚丙烯酰胺凝胶电泳和蛋白免疫印迹技术分析籼稻和粳稻热稳定蛋白的表达差异.探讨利用特异蛋白来建立籼粳稻的鉴别方法.结果表明,在籼粳稻品种之间存在热稳定蛋白的差异表达,尤其在分子量约42～47kD范围.其中45.2kD条带(Band I)和46.5 kD(Band Ⅱ)条带为典型粳稻特异蛋白(Os03g0168100)的标志带,42.0kD条带(Band Ⅲ)为典型籼稻特异蛋白(OsI_10172)的标志带.以这3条带作为鏊别方法,并与程氏指数法进行比较,典型籼稻和粳稻的一致度分别为80.0%和86.4%,表明热稳定蛋白标志带法在一定程度上可用于鉴别典型的籼稻和粳稻.     

人工老化对老芒麦种子活力和生理生化变化的影响

以4份老芒麦种子为试验材料,利用100%RH高温高湿老化法(AA)对老芒麦种子进行不同梯度的老化处理,探讨老化后老芒麦种子活力和种子内部生理生化的变化。结果表明:随着老化程度的加深,4份老芒麦种子的各项活力指标均出现明显下降趋势;生理生化指标中,种子浸出液电导率出现先下降后缓慢上升、可溶性糖含量呈现先减少后增加的现象,而丙二醛含量则出现整体一致的上升趋势,POD活性变化无规则,SOD活性出现先增加后减小的现象,CAT活性则发生整体下降的趋势。本研究结果将为老芒麦种质资源的保护和遗传完整性的研究提供基础理论依据。     

高温处理对转基因作物种子Bt蛋白含量的影响北大核心CSCD

转基因作物食品安全问题日益受到社会关注,作物种子作为食物的重要来源之一,转基因作物种子的安全性研究更有待加强。分别利用转Bt基因棉花和玉米品种及非转基因品种种子进行高温处理(55-130℃),并用双抗夹心酶免疫检测方法(ELISA)研究其中的外源Bt蛋白的动态变化过程。结果表明,不同作物种子中的Bt蛋白对高温的耐受程度不同,棉花种子中Bt蛋白较玉米种子耐高温能力强;高温处理条件下,种子中的Bt蛋白在10 min内含量显著下降,其后降解缓慢;130℃以上的高温处理条件下,玉米和棉花种子中的外源Bt蛋白均能显著降解。     

人工老化对春小麦种子萌发及根尖超微结构的影响

选取‘新春6号’小麦种子为材料,进行不同时间的高温高湿人工加速老化处理,考察发芽率、幼苗根长、幼苗鲜重、幼苗干重等指标,并用扫描电镜和透射电镜观察其种子根尖表皮形态及细胞结构变化,明确不同老化处理对春小麦种子萌发及根尖细胞超微结构的影响。结果表明,随着种子劣变程度加深(老化处理时间的延长),春小麦种子发芽率、发芽势、发芽指数、幼苗根长、鲜重、干重等相关活力指标急剧下降,而平均发芽时间持续增加,且以老化处理4d为种子活力下降的转折点。种子萌发72h后,胚根尖表皮结构的扫描电镜观察发现,随着老化程度加深,胚根尖表皮皱缩逐渐加深、紧凑,最终致使表皮破裂,内部细胞裸露;透射电镜观察则显示,老化处理越久,根尖细胞中细胞器溶解程度越明显,最终导致整个细胞形成空腔。可见,根尖表皮形态、内部细胞结构上的改变可能是引起种子逐步丧失活力的原因。     

甜高梁种子人工老化过程中活性氧清除酶活性的变化

研究甜高粱品种'意达利'种子人工老化过程中种子活力与活性氧清除酶活性之间关系的结果表明:随着老化处理(100%RH,43℃)的时间进程,种子的萌发率、活力指数以及由存活种子形成的幼苗鲜重下降,相对渗漏率增加,丙二醛含量下降.超氧物歧化酶、抗坏血酸过氧化物酶、过氧化氢酶、谷胱甘肽还原酶和脱氢抗坏血酸还原酶的活性在人工老化初期增加,而后下降.据此认为活性氧清除酶活性下降可能是甜高粱种子活力丧失的原因之一.     

不结球白菜种子活力及抗氧化特性在人工老化过程中的变化

以不结球白菜品种‘寒笑’种子为材料,研究高温(42℃)高湿(相对湿度100%)人工老化处理过程中种子活力及抗氧化相关特性的变化。结果显示:不结球白菜种子的发芽率、发芽势、发芽指数和活力指数随老化处理时间的延长而逐渐下降,不正常苗率逐渐增加;种子的超氧阴离子(O2.-)产生速率先增高后降低,过氧化氢(H2O2)含量逐渐增加,脂氧合酶(LOX)活性和丙二醛(MDA)含量下降,种子浸出液相对电导率增加;种子超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽还原酶(GR)活性随老化处理时间的延长逐渐下降,抗坏血酸过氧化物酶(APX)和谷胱甘肽过氧化物酶(GPX)活性在老化处理初期(老化3 d前)均增加,APX活性随后降低,GPX活性无显著变化;种子抗坏血酸(AsA)和还原型谷胱甘肽(GSH)含量在老化处理1 d后即显著降低并持续保持较低水平。研究表明,不结球白菜种子在人工加速老化过程中其抗氧化系统代谢紊乱并造成活性氧累积伤害,这可能是引起不结球白菜种子老化的重要原因。     

高温加速老化过程中甜菜种子线粒体小分

甜菜(BetavulgarisLcv.Loke)种子的人工老化(30℃,100％RH)导致种子的干重、生活力和活力逐渐丧失,线粒体的呼吸速率、细胞色素c氧化酶和苹果酸脱氢酶的活性下降。线粒体蛋白质随着种子老化而显著变化,小分子量热休克蛋白(LMWHSP)22从老化的0d到30d增加,然后从30d到90d下降。可以认为甜菜种子人工老化过程中线粒体活性和LMWHSP22的含量变化与种子活力密切相关。     

高温加速老化过程中甜菜种子线粒体小分 子量热激蛋白22的变化

甜菜(BetavulgarisLcv.Loke)种子的人工老化(30℃,100％RH)导致种子的干重、生活力和活力逐渐丧失,线粒体的呼吸速率、细胞色素c氧化酶和苹果酸脱氢酶的活性下降。线粒体蛋白质随着种子老化而显著变化,小分子量热休克蛋白(LMWHSP)22从老化的0d到30d增加,然后从30d到90d下降。可以认为甜菜种子人工老化过程中线粒体活性和LMWHSP22的含量变化与种子活力密切相关。     

逆境条件下小麦种子活力与种子萌发相关酶活性及其基因表达的关系

分析不同基因型小麦品种逆境萌发过程中种子萌发相关酶活性及基因表达差异,明确在逆境条件下,种子活力与种子萌发相关酶活性及基因表达量的关系.通过标准发芽试验和逆境(冷浸、人工老化、干旱胁迫)发芽试验,测定4个小麦品种种子活力、萌发过程中可溶性总糖和可溶性蛋白含量、α-淀粉酶活性、半胱氨酸蛋白酶活性及相关基因表达量.结果表明:干旱、人工老化和冷浸胁迫3种逆境对种子活力都有一定影响.不同萌发条件下,可溶性总糖含量呈现先小幅度升高后小幅度降低再迅速升高的趋势;而可溶性蛋白含量随着萌发时间的延长呈现逐渐下降的趋势.α-淀粉酶活性整体呈现逐渐升高的趋势,但在冷浸胁迫处理后,豫农949和轮选061的α-淀粉酶活性在萌发60 h后出现下降.半胱氨酸蛋白酶活性整体呈先降低后升高的趋势,但在干旱胁迫条件下,豫农949、豫麦49-198和轮选061的半胱氨酸蛋白酶活性呈现先升高后降低再升高的趋势.不同逆境萌发条件下,α-AMY(α-淀粉酶基因)表达量整体呈先上升后下降的趋势.冷浸胁迫处理后,轮选061的α-AMY表达量高于对照,在其他逆境萌发条件下,4个品种的α-AMY表达量均低于对照;人工老化处理后,长4738的CP(半胱氨酸蛋白酶基因)表达量与对照差异不显著,在其他逆境萌发条件下,4个品种的CP表达量均高于对照.种子萌发期间,不同萌发条件下α-淀粉酶和半胱氨酸蛋白酶活性与其基因表达并没有直接关系,α-淀粉酶活性与可溶性总糖含量达到显著正相关,半胱氨酸蛋白酶活性与可溶性蛋白含量的相关性不显著.在标准发芽条件下,α-淀粉酶活性与活力指数呈显著正相关,而在逆境萌发过程中,其相关性不显著.冷浸胁迫处理后,半胱氨酸蛋白酶活性与活力指数呈显著正相关,但在标准发芽、干旱胁迫、人工老化处理后,其相关性不显著.     

Comparative studies of genetic diversity in kenaf (Hibiscus cannabinus L.) varieties based on analysis of agronomic and RAPD data

Kenaf (Hibiscus cannabinus L.) is a fiber crop classified in the genus Hibiscus (Malvaceae), and has a great potential for its multipurpose utilization, in addition to its traditional usage. Varietal identification of kenaf is always problematic and knowledge on genetic diversity of kenaf varieties is also limited, which significantly hindered our effective utilization and conservation of the valuable kenaf germplasm. In order to find a proper method for identifying kenaf varieties and studying their variation, morpho-agronomic characters and random amplified polymorphic DNA (RAPD) markers were analyzed among 14 kenaf varieties commonly used in Japan. Data from morphological analysis showed that the included kenaf varieties could be divided into three major groups. The characters, such as middle stem diameter, whole stalk weight, and days to 50% flowering, are highly responsible for the variation of the kenaf varieties, but it is difficult to identify individual varieties merely by the morpho-agronomic characters. On the other hand, clearly separation of the kenaf varieties was achieved based on the RAPD variation patterns. Genetic relationship of the kenaf varieties can also be traced through the analysis of RAPD and morph-agronomic variation. It is concluded from the present study that RAPD analysis is an effective tool in identifying of kenaf varieties and determining their genetic relationships, particularly when combined with the analysis of morpho-agronomic characters.     

红麻(Hibiscus cannabinus L.)种质资源的鉴定与分类研究

对300份野生红麻和337份栽培红麻的形态学鉴定发现,红麻的茎、叶、花、果等植物学性状的类型多样,差异明显,遗传多样性十分丰富。野生红麻和栽培红麻一样,染色体数为2n=36,同属红麻种（Hibiscus cannabinus L．）;核型有2n=36=32M＋2SM＋2M（SAT）、2n=36=34M＋2M（SAT）和2n=36=32M＋2ST＋2M（SAT）等;综合形态学研究结果,将野生红麻分为普通、多毛、螺旋花和扁圆果4种类型,其中普通型又可分为3个亚类,普通型与栽培红麻亲缘关系最近,并发现红麻存在亚种。栽培红麻可分为9个变种17个类型,其中4个为新发现变种,7个为新发现类型。     

Variety Trial and Pyrolysis Potential of Kenaf Grown in Midwest United States

Kenaf (Hibiscus cannabinus L.) has potential as an annual herbaceous biomass feedstock. It is not typically grown in the American Midwest; however, kenaf may be attractive as an alternative crop for Iowa and the Corn Belt. In this study, seven kenaf varieties were grown in Iowa and evaluated for their productivity. More specifically, our research questions were the following: (1) how do kenaf varieties perform in Iowa for yield? (2) How does fiber morphology and quality differ among varieties and among core and bast fiber? And (3) What potential does kenaf (bast and core) have for producing fuel using fast pyrolysis? Tainung 2, one of the varieties, reached the best yield in Central Iowa over multiple years. Bast kenaf contained 8 % more cellulose and 23 % less hemicellulose than the core but it varied among varieties. Also, regardless of variety, core was composed of 40 % more lignin than bast. Core was found to have higher potential for fast pyrolysis than the bast but its potential was variety-dependent. Overall, kenaf could be grown to diversify Iowa agriculture and provide alternative feedstock to the biofuel industry.     

Genetic Diversity of Kenaf (Hibiscus cannabinus) Evaluated by Inter-Simple Sequence Repeat (ISSR)

Understanding genetic diversity is very useful for scientific utilization for breeding. In this study, we estimated the genetic distances in a panel of 84 kenaf accessions collected from 26 countries and regions using ISSR markers. The results of UPGMA analysis showed that kenaf germplasm had abundant genetic variation, with genetic dissimilarity coefficients ranging from 0.01 to 0.62. The in-group dissimilarity coefficient (0.29) was observed in 84 kenaf accessions, and all the accessions could be divided into three groups: cultivars (L_1–1), relatively wild species (L_1–2 and L_1–3), and wild species (the others). Further in-group analysis in group L_1–1 (0.19) revealed that the kenaf cultivars could be divided into five subgroups with distinct regional characteristics. It is imperative that genes be exchanged among all kinds of tested varieties from different origins. The results provide a useful basis for kenaf germplasm research and breeding.     

Genetic diversity and phylogenetic relationship of kenaf (Hibiscus cannabinus L.) accessions evaluated by SRAP and ISSR

Understanding genetic diversity and phylogenetic relationships is useful for plant breeding. In this study, we assessed the genetic diversity in a panel of 84 accessions of kenaf from 26 countries using SRAP and ISSR markers. The kenaf accessions could be divided into L1 (60 cultivated varieties) and L2 (24 wild accessions) at the level of 0.145 genetic dissimilarity coefficient by UPGMA. The L₂ group was further divided into two subgroups (16 relative-wide and 9 origin wide accessions) at the level of 0.207 genetic dissimilarity. Out of the 9 wild accessions in the L₂ group, 6 were from Tanzania and the remaining 3 lines were from Kenya. These results suggest that the center of origin for kenaf might be Tanzania and Kenya.     

Protein kinases in brown adipose tissue of developing rats. II. two soluble kinase activities and their affinities for nucleotide and protein substrates.

A heat-stable, soluble component of brown adipose tissue from newborn rats was found to be readily phosphorylated by protein kinase of the same subcellular fraction. The concentration of this component in brown fat decreased with the age of the animals. A boiled crude microsomal preparation from rat liver was also phosphorylated by brown fat protein kinase. The GTP-linked phosphorylation of the endogenous heat-stable protein was not stimulated by ATP (in contrast to phosphorylation of histone). The maximum velocity of phosphorylation achieved with GTP was about 2.5 times higher than that with ATP as nucleotide substrate. This difference was not due to ATPase activity in the assay. With histone as the protein acceptor both activities were the same. The affinity of protein kinase(s) for ATP was lower with the endogenous heat-stable brown-fat protein and with boiled microsomes (Km of 0.21 mM and 0.17 mM, respectively) than with histone (Km of 0.05 M). No detectable ATPase activity was present in either acceptor protein. It is concluded that the 100 000 times g supernatant fraction from brown fat of infant rats contains two protein kinase activities. One preferentially uses ATP and histone as substrates and the other uses endogenous heat-stable soluble proteins and either ATP or GTP.     

红麻微卫星DNA标记指纹图谱的构建

DNA指纹图谱对新品种选育、种质资源保存和管理具有重要的意义。然而,利用SSR标记构建红麻DNA指纹图谱的研究仍十分有限。在本研究中,利用课题组开发并筛选出的131对SSR引物,分析不同来源的96份红麻种质资源,包括红麻品种审定的区试对照品种福红952。结果表明,131对引物共扩增出375条带,平均每对引物扩增出2.6条带。以遗传相似系数0.614为切割线时,可以分为2个类群,52个为类群P1,44个为类群P2;以遗传相似系数0.710做切割线,可分为5个亚群。利用这131对引物标记所得的数据成功绘制了一份85个品种独特的指纹图谱,其中福红952可被HcEMS238引物特异识别。其他11份因存在遗传相似性高的现象,未被识别。上述结果为红麻品种的真实性鉴定及遗传多样性分析提供依据。     

A comparative study of the atp9 gene between a cytoplasmic male sterile line and its maintainer line and further development of a molecular marker specific for male sterile cytoplasm in kenaf (Hibiscus cannabinus L.)

mtDNA was isolated from cytoplasmic male sterility (CMS) line P3A and its maintainer P3B of kenaf (Hibiscus cannabinus L.). The atp9 gene and its two flanking sequences were obtained using homology cloning and high-efficiency thermal asymmetric interlaced PCR methods. The coding sequences showed only two base pairs difference between the CMS and its maintainer, and shared a homology of over 87 % with atp9 genes from other species in GenBank. However, when comparing the flanking sequences, a 47-bp deletion was characterized at the 3′ flanking sequence of atp9 in the CMS line. Quantitative PCR analysis indicated that the expression level of atp9 in the CMS line was 0.937-fold that of its maintainer. Furthermore, the respiratory rate of anthers in the CMS line was markedly lower than that of its maintainer. The results indicated that the 47-bp deletion at the 3′ flanking sequence of atp9 and/or down-regulated expression of the atp9 gene in the CMS line might be closely related to CMS in kenaf. To confirm whether the 47-bp deletion was specific to cytoplasm of male sterile lines, another 21 varieties were used for further analysis. The results showed that the 47-bp deletion was specific to male sterile cytoplasm (MSC) of kenaf. Based on these, a specific molecular marker was developed to distinguish the MSC from male fertile cytoplasm of kenaf.     

生菜矿质养分特性的品种差异比较研究

为了探明生菜不同品种类型之间的矿质养分特性差异,本实验采用深水栽培方式,比较研究了玻璃生菜、油麦菜和意大利生菜3个品种在不同生长时期地上部Fe、Zn、Ca含量的差异,以及根际缺氧处理对3个品种地上部Fe、Zn、Ca含量的影响.结果表明,随着植株的生长,3个品种地上部的Fe、Zn、Ca含量均呈上升趋势,到出苗后50 d达到最高水平,随后稍有下降.3个品种地上部3种矿质养分含量由高到低的顺序依次是：意大利生菜、玻璃生莱、油麦菜.但3个品种地上部的生长速率的大小顺序却相反.根际缺氧处理（以正常通气栽培植株作对照）均使3个品种地上部的Fe、Zn、Ca含量下降,其中意大利生菜所受的影响最大,其地上部的Fe、Zn、Ca含量分别比对照降低18.04%、12.70%、17.56%;油麦菜的影响最小,其地上部Fe、Zn、Ca含量的降低率不及意大利生菜的一半;玻璃生菜地上部Fe、Ca含量受影响比较大,分别比对照降低10.54%和14.79%,但Zn含量的影响比较小.     

Expression and characterization of the extracellular domain of guanylyl cyclase C from a baculovirus and Sf21 insect cells

Guanylyl cyclase (GC)-C, a single-transmembrane receptor protein for heat-stable enterotoxin, guanylin, and uroguanylin, and its N-terminal extracellular domain were prepared at a high level of expression from a system constructed of Sf21 insect cells and recombinant baculovirus. The recombinant GC-C, containing the complete sequence, retained its binding affinity to heat-stable enterotoxin with a KD value (6.2 x 10(-10) M) and cyclase catalytic activity at a level similar to those of GC-C expressed in mammalian cell lines, such as COS-7. The N-terminal extracellular domain was prepared in a form which contained the hexahistidine tail at its C-terminus and was purified as a homogenous protein by Con A and Ni-chelating affinity chromatography from the culture medium of the insect cells. The purified N-terminal extracellular domain of GC-C exhibited the high (KD = 4 x 10(-10) M) and low (KD = 7 x 10(-8) M) affinity sites in binding to heat-stable enterotoxin. These results clearly indicate that the N-terminal extracellular domain of GC-C possesses the same biochemical characteristics as the complete GC-C protein even in the membrane-free form. Moreover, the extracellular domain is able to form an oligomer in a ligand-dependent manner, suggesting that the N-terminal extracellular domains interact with one another in binding to ligands.