SDS-PAGE、同工酶等电聚焦和RAPD标记鉴定两系杂交水稻F1种子纯度对比研究

目的：探讨利用种子贮藏蛋白SDS—PAGE电泳、酯酶同工酶超薄等电聚焦电泳和RAPD分子标记鉴定五个两系杂交水稻组合F_1种子纯度的可行性。方法和结果：利用SDS—PAGE电泳技术未能找到所试五个组合各自的父本特征蛋白质带;利用酯酶同工酶超薄等电聚焦电泳技术和RAPD分子标记可找到所试五个组合的父母本特征酶带和RAPD标记,但酯酶同工酶的多态性同时也受种子萌发时间的影响。酯酶同工酶超薄等电聚焦电泳和RAPD分子标记可用于所试五个两系杂交水稻组合F_1种子纯度的鉴定。     

用等电聚焦技术鉴定杂交稻华优桂99的种子纯度

利用等电聚焦技术对杂交稻华优桂99及其亲本种子蛋白进行了分析,发现有一对亲本互补谱带.应用此种互补谱带,对华优桂99的种子纯度进行了鉴定,与田间小区种植鉴定的结果比较差异较小.表明这一技术在华优桂99种子纯度的鉴定中是可行的.     

三种分析水稻F1代幼苗生长过程中酯酶同工酶变化的电泳方法比较

利用聚酯胶片作为凝胶支持物的超薄等电聚焦电泳(UTLIEF)研究水稻F1种子萌发过程中的酯酶同工酶多态性变化的结果表明,UTLIEF相比非变性不连续聚丙烯酰胺电泳(native-PAGE)和常规等电聚焦电泳(IEF)得到的酯酶同工酶图谱更清晰,酶带数目和强弱的多态性变化更高,用pH 2～9的两性电解质时等电聚焦的电泳效果较好.     

一种改良的醋酸纤维素膜血清蛋白等电聚焦电泳方法

目前在等电聚焦电泳技术中,应用最多的是聚丙烯酰胺凝胶平板薄层等电聚焦。该法具有分辨率高的优点,但需采用高纯度试剂,价格昂贵,而且超薄层制板(≤0.5mm)技术要求也较高,这些条件限制了它的推广应用。利用醋酸纤维素膜进行等电聚焦,则可弥补上述缺点。但众所周知,醋酸纤维素膜存在较强     

杂交水稻汕优63杂种纯度的RAPD鉴定

使用纯度低或真实性差的杂交种子会给农业生产造成极大损失。然而杂交种子的纯度无法单纯从种子形态上鉴别。本文首次采用RAPD特异扩增谱带做为分子标记对水稻杂交种纯度进行了鉴定。汕优63杂交水稻种子由中国水稻研究所从某制种单位随机取样获得。珍汕97A不育系和明恢63恢复系等品种由于中国水稻研究所提供。在做形态观察的同时,分别取上述三种材料叶片制备DNA作为初始模板DNA。对300个RAPD随机引物,经过三次多态性初筛复筛,发现随机引物P18可发稳定地扩增出一条来源于父本明恢63的0．8kb的特异条带。用P18引物对100株汕优63杂交单株进行RAPD扩增,其中83个获得了0．8kb特异条带（Fig.1upper)。分子交杂证明其结果是可靠的（Fig,1Lower)。以RAPD扩增结果为依据,对这100株材料进行植株形态比较,发现那17个不能扩增出特异条带的单株均为假杂种。说明该制种单位汕优63的假种率为17％。大大超过部的有关规定。应对其原因和后果进行追究。用P18引物对18种分别为汕粳及其中间型的常用稻种进行PAPD扩增鉴定。除明恢63的亲本－圭630和中间型Pecos,Aus373具有0．8kb的特异条带（Fig.2）以外,其余均没有。是否明恢63与这两个中间型有起源关系,尚有待进一步验证。但至少说明：PAPD扩增可用于鉴定水稻品种田间串粉造成的假杂种。理论上,两个不同品种的杂交F1代的PAPD扩增谱带应为两种类型：与双亲同型和互补型。远大实际上,由于PAPD引物对不同DNA自然的竞争扩增效应等因素影响,常常使扩增量少的一些DNA片段在电泳谱带上表现不出来,而表现为单一亲本型谱带。这是引物P18之所以只有一条父本型条带的。多数情况下,这种单一新本型谱带也可以作为鉴定杂种纯 依据,但是要参照诸如形态特征等其它指标,区分具有同样大小（假定存在0．8kb的其怂u）特异条带的假杂种。本实验室正在继续筛选来源于母本的单一亲本型或互补型谱带的随机引物,立图建立起仅用PAPD扩增检测即可确诊的简便方法。     

单克隆抗体的等电聚焦

本文采用聚丙烯酰胺凝胶等电聚焦和Phast-system仪等电聚焦测定12份HBsAg McAb IgG的等电点,并对两种方法测定的误差和优缺点进行了讨论。     

特优系列杂交稻种子纯度的鉴定

用等电聚焦技术(IEF)对特优系列品种及其亲本种子蛋白进行了分析,2-氯乙醇不能提取F1代的亲本互补蛋白带,而用40%的2-氯乙醇与15%的巯基乙醇混合液则能有效提取F1代亲本互补带.采用此种互补谱带,可鉴定"特优524"和"特优721"品种的种子样品,其结果与田间小区种植鉴定的差异较小.     

全柱成像毛细管等电聚焦电泳测定艾塞那肽等电点

目的:采用新一代全柱成像毛细管等电聚焦电泳技术(CIEF-WCID)测定艾塞那肽等电点。方法:采用互补性金属氧化物半导体成像技术对样品等电聚焦过程进行实时记录,根据适宜的marker计算得到艾塞那肽的等电点,并对方法的准确度与重复性进行考察。结果:测得艾塞那肽等电点为5.46,与凝胶电泳结果基本一致,相对标准偏差为0.11%。CIEF-WCID方法快速准确,相对误差小于2.5%,重复性良好。结论:CIEF-WCID作为一种新的技术手段可用于艾塞那肽等电点的分析,方法快速、准确、重复性好,可为多肽的质量控制提供一种可靠的分析方法。     

目前最高分辨率的电泳──固相pH梯度等电聚焦

固相pH梯度等电聚焦是国际上80年代的新型电泳技术.利用一系列具有弱酸和弱碱性质的丙烯酰胺衍生物滴定时,在滴定终点附近形成的pH梯度并参与丙烯酰胺的共价聚合,从而形成固定的不随环境电场等条件变化的pH梯度,该方法具有比传统载体两性电解质等电聚焦更高的分辨率、更大的上样量.可用于分析和制备相近pI的蛋白质,多肽等.     

薄层凝胶等电聚焦电泳的快速简便制胶方法

在薄层凝胶等电聚焦电泳技术中,制胶可谓是关键的一步。如果制胶失败,将造成时间上的浪费和经济上的损失(凝胶中所用的两性电解质载体——安福林的价格较昂贵)。如果制成的凝胶薄厚不匀或无支撑物,将会直接影响实验结果或给实验操作带来麻烦。我们针对这种情况,结合实验教学,摸索出一种快速、简便而效果较好的制胶方法。现简要介绍如下     

Seed Number, Seed Size and Seed Diversity in Washington Lupin (Lupinus polyphyllus Lindl.)

Over 20 000 seeds of the Washington lupin (Lupinus polyphyllusLindl.) were examined and measured in an experiment carriedout over a 10 year period (1989–1999). Four differentgroups of Washington lupin seeds were found: dark, patterned,grey and light seeds. During the 10 year experiment, the totalaverage number of seeds per plant decreased from 2654 (1990)to 1220 (1999), there was a slight decrease in seed weight perplant and an increase in the average weight per seed. Therewas a clear seed size/number trade-off at the intraspecificlevel. The relative proportions of each seed group also changedwith patterned seeds becoming dominant (50% at the beginningand 90% at the end of the experiment), grey seeds remained constant(constituting 10% of the total seeds) and the proportion ofboth dark (33% at the beginning and 5% at the end) and light(10% at the beginning and 5% at the end of the experiment) seedsdecreased. Six hundred and sixty nine seeds were found to havea different testa ornamentation; they were distributed amongthe different groups as follows: 48% patterned seeds, 29% darkseeds, 12% grey seeds and 11% light seeds. There were no significantdifferences in the physical dimensions of the ornamented seedscompared with all other seeds. The results suggest that theWashington lupin is a mixture of different species and botanicalforms; this is discussed in relation to possible selection pressuresto produce both smaller and larger seeds. The possibility thatchanges in testa ornamentation are influenced by genes controllingthe synthesis of the seed coat pigment is discussed. Copyright2001 Annals of Botany Company Lupinus polyphyllus Lindl., lupine, plant biotechnology, plant parameters, selection pressure, seed size/number trade-off, testa ornamentation     

Seed flavonoids of thePodalyrieae andLiparieae (Fabaceae)

A study of the phenolic compounds of the closely related papilionoid tribes,Podalyrieae andLiparieae, proved that the flavonoid patterns of hydrolysed seed extracts are remarkably conservative. Butin (7, 3, 4-trihydroxyflavanone), 3-hydroxydaidzein (7, 3, 4-trihydroxyisoflavone), vicenin-2 (6, 8-di--D-glucopyranosyl-5, 7, 4-trihydroxyflavone) and orobol (5, 7, 3, 4-tetrahydroxyisoflavone) were isolated and identified as the major flavonoids. The seeds ofAmphithalea, Coelidium, Liparia, Xiphotheca, Calpurnia, Stirtonanthus andPodalyria accumulated three isoflavone O-glycosides that yielded 3-hydroxydaidzein on hydrolysis. In contrast,Virgilia contained a unique combination of vicenin-2 and orobol. Vicenin-2 was also present inCalpurnia as a major compound, butStirtonanthus insignis was the only other species studied that contained orobol (in trace amounts only). Butein, a chalcone, was reported byHarborne from the seed ofCyclopia subternata. This compound's flavanone analog, butin, was the principal component inCyclopia. A cladistic analysis, using flavonoid, alkaloid and morphological data, showed that the seed flavonoids of thePodalyrieae andLiparieae behave rather poorly as cladistic characters. They are, however, of considerable taxonomic value at the tribal level favouring the opinion that the two tribes should be combined. The apparent absence of flavonoids in the seed ofHypocalyptus supports the suggestion that it should be excluded from theLiparieae. Flavonoids also show that theArgyrolobium-group is very different from the tribeCrotalarieae and support the recent transfer of this group to the tribeGenisteae.     

Seed development ofAbelmoschus esculentus (Malvaceae)

The endosperm is nuclear, cell wall initiation starts 5 days after pollination. During early stages endosperm nuclei exhibit synchrony in their division. Embryogeny is of the Asterad type. A7-to 10-celled suspensor persists up to the dicot stage of the embryo. Both integuments contribute towards formation of the seed coat. 30 days after pollination seeds become mature. Their endosperm is scanty and persists as a thin layer between the folds of the cotyledons. Nucellus remnants are present towards the funicular side.     

Seed dispersal characteristics of Brassavola nodosa (Orchidaceae)

We used mathematical models for wind-dispersed seeds and wind-tunnel experiments to predict modal seed dispersal distance of the Neotropical orchid Brassavola nodosa under conditions approximating those found in its natural habitat: mangrove islands in Belize, Central America. Key variables in a simple ballistic model for predicting modal dispersal distance (xm) of an individual seed include: height of release (h); free-stream velocity (Uc); and terminal velocity of the seed (Ut): xm = h Uc/Ut. Modal dispersal distance of dust-like orchid seeds were predicted adequately by this ballistic model at low wind velocities and low release heights, but it underestimated the increasing importance of turbulence at higher wind velocities and greater release heights. We estimated the magnitude and relative importance of one measure of turbulence, vertical mixing velocity (W*), on xm in wind tunnel experiments. Our estimates of W* were within the same order of magnitude as those found for other small dust-like seeds and pollen. In high turbulence conditions, incorporation of vertical mixing velocity effects into the ballistic model of seed dispersal overestimated modal seed dispersal distances.     

螺序草属(茜草科)的种子形态

对螺序草属(Spiradiclis B1、)16种2变型共18个样品的种子形态特征的初步研究表明,本属植物种子为小型（0．2—0、3mm)或中型(0．3—2mm),整体形状不太规则,表面纹饰为蜂窝状,种脐稍突出。种子的外壁由内外2层种皮组成。依据表面突起分布方式,螺序草属的种子大致分为2种类型：(1)平周壁微下陷,疣突均匀分布于其上,如大叶螺序草、螺序草、尖叶螺序草、柳叶螺序草、峨嵋螺序草和龙州螺序草;(2)平周壁下陷成穴状,突起分布在穴边缘,如红花螺序草、心叶螺序草、两广螺序草、广东螺序草、海南螺序草、宽昭螺序草、疏花螺序草、多枝螺序草、罗氏螺序草、小叶螺序草、石生螺序草和紫花螺序草。螺序草属的种子形态特征对属下类群的划分具有重要的参考价值。     

Seed morphology in Southern AfricanOrchidoideae (Orchidaceae)

The seed morphology of 151 species of Southern AfricanOrchidoideae (Orchideae andDiseae; sensuDressler 1981) was studied by means of scanning electron microscopy. Two different seed types were found. (1) In the majority of species the seeds are minute and fusiform. The seed coat is made up of comparatively few concave and elongate testa cells with straight or slightly undulate and generally unthickened anticlinal cell walls. The seed type was here termed Satyrium-type. While most species are very similar in the ornamentation of the periclinal walls of their seed coat, considerable variation was found inHolothrix where two distinct groups can be recognized in this respect. (2) A remarkably different seed type was observed inDisa uniflora and three apparently closely related species (Disa uniflora-type), where large balloon-like seeds occur. Their seed coat consists of convex cells with undulate anticlinal walls. It is suggested that this seed type is a derived condition and has evolved in adaptation to the specialized habitat alongside streams. The possibility of hydrochory in these four species is briefly discussed.     

Seed ontogeny in Adesmia securigerifolia (Fabaceae-Adesmieae)

A study of the reproductive processes of Adesmia securigerifolia from bud to mature seed was carried out by means of field observations and the paraffin technique. Observations revealed the following new contributions to the study of legume embryology: 1) after fertilization, a small nucellar haustorium, or micropylar nucellar beak, was observed for the first time, originating from two obliterating nucellar cells that extended outwards. Their globose distal end comes in contact with the internal carpel wall, while the wedge shaped base stretches into the micropyle; a suspensor consisting of five or more cells - the two basal cells are large and falcate and fit into the micropylar pore - coexists with the undivided polar nuclei thus showing that endosperm formation begins after zygote division; 2) at the young embryo stage, a sac- shaped nuclear haustorium, formed by the endosperm, adjoins the outer integument and is not connected to the chalaza, or any vascular element; at the hilar level, a nucellar projection is formed in connection with the haustorial coenocytic endosperm. This projection persists up to the mature seed stage when it starts to degenerate, after performing another linking with the embryo nutrition system; 3) at the mature seed stage, the seed coat evolving from the outer integument has a single macrosclereid layer, though inclusions in the cell vacuoles simulate the presence of more layers and/or transverse walls. The lens, a hypodermal layer of osteosclereids (hour-glass cells), and the astrosclereids are also described.     

Seed abortion in Pongamia pinnata (Fabaceae)

In Pongamia pinnata only one of the two ovules develops into a seed in most of the pods. Since pollen was not found to be limiting and reduced fertilization could not completely explain the observed frequency of seed abortion, it implied an effect of postfertilization factors. Aqueous extracts of developing seeds and maternal tissue (placenta) did not influence abortion in vitro, suggesting that abortion may not be mediated by a chemical. Experimental uptake of ¹⁴C sucrose in vitro indicated that both the stigmatic and the peduncular seed have similar inherent capacities of drawing resources, but the peduncular seed is deprived of resources in the presence of the stigmatic seed. This deprivation of the peduncular seed could be offset by supplying an excess of hormones leading to the subsequent formation of two seeds in a pod. The prevalence of single-seeded pods in P. pinnata seems therefore to be a result of competition between the two seeds for maternal resources. The evolutionary significance of single-seeded pods in P. pinnata is discussed with respect to possible dispersal advantage enjoyed by such pods.     

Verticillium dahliae (Kleb.) Infecting Pumpkin Seed

This study investigated the natural occurrence of Verticillium dahliae (Kleb.) infection in pumpkin (Cucurbita pepo L.) seed. The mean incidence of infection was found to be 21.0%. Isolates recovered from seeds were pathogenic to pumpkin (cultivar ‘Jamaican squash’). Surface sterilization by immersion in 0.6% sodium hypochlorite for 20 min eradicated V. dahliae from infected pumpkin seeds without affecting germinability. Plating of seed components revealed that the fungus was present in the seed coat but not in the embryo or cotyledons. In a growing‐on test, 25% of 6‐week‐old plants grown from untreated seeds were infected. Germination and production of normal seedlings were unaffected by V. dahliae infection of seeds. Verticillium dahliae in pumpkin seed was found to be external and transmissible to plants. The findings of this study are important in devising disease control strategies.     

Seed coat anatomy ofCeratozamia mexicana (Cycadales)

The seed coat furnishes protection with a thick cuticle, tannin cells, mucilage, and a hard sclerotesta. The external layer of the seed coat is a sarcotesta; a thick cuticle covers the external walls of its epidermal cells. This epidermis bears stomates and, in the early stages, trichomes. The subepidermal cells have druses. Starch grains are abundant in the sarcotesta from June through August, but they disappear during dispersal in September. The parenchyma is interrupted by mucilage canals lined by epithelial cells. Tannin cells are found in the sarcotesta, sclerotesta, and pachychalaza. Ten sectors of an areole in the sclerotesta around the micropyle may correspond to the tips of the integumentary segments in some fossil plants, such asGenomosperma kidstonii.

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Resumen  La cubierta seminal le imparte protección a la semilla mediante una cutícula gruesa, células de taninos, mucílago, y una esclerotesta dura. La capa externa de la cubierta seminal es la sarcotesta; una cutícula gruesa cubre las paredes externas de las células epidérmicas. Esta epidermis presenta estomas y, en etapas tempranas, tricomas. Cuando la esclerotesta se endurece, los tricomas desaparecen. Las células subepidérmicas presentan drusas. En la sarcotesta los granos de almidón son abundantes desde junio hasta agosto, pero desaparecen durante la dispersión en septiembre. El parénquima se interrumpe por canales de mucílago forrados por células epiteliales. Las células de taninos se encuentran en la sarcotesta, esclerotesta, y paquicálaza. Diez sectores de una areola en la esclerotesta alrededor del micrópilo podrían corresponder a los apéndices del tegumento en algunas plantas fósiles, comoGenomosperma kidstonii.