无籽沙糖桔WUBE1基因的功能验证

为了解泛素活化酶E1基因(UBE1)在无籽沙糖桔自交不亲和反应中的作用,通过根癌农杆菌介导法将来源于自交不亲和无籽沙糖桔(Citrus reticulata ‘Wuzishatangju’) WUBE1基因转化烟草(Nicotiana tabacum)。结果表明,外源基因WUBE1已导入烟草基因组中并得到表达。转WUBE1基因的自交授粉组合花粉管在生长过程中,部分花粉管出现停止生长的现象,到达花柱基部的花粉管数量少于异交授粉和野生型自交组合。转WUBE1基因烟草的花粉生活力、发芽率、自交和异交后每个果荚中的种子数与野生型烟草无显著差异。这表明单一的WUBE1基因不能调控无籽沙糖桔自交不亲和反应,很可能是通过Ub/26S途径参与了无籽沙糖桔自交不亲和反应。     

四倍体青花菜低稔性胚胎学研究

以二、四倍体青花菜为材料,在现蕾期分别套袋自交,对四倍体与二倍体的胚囊结构、受精以及受精后胚胎发育过程进行比较研究.结果表明:四倍体青花菜胚囊发育完整,与二倍体相似.四倍体花粉萌发率较二倍体低,花粉管生长较慢且不整齐.四倍体和二倍体青花菜自交均能正常受精,四倍体青花菜在受精后胚胎发育的进程上均较其二倍体滞后约2 d,且胚胎发育在进度上不一致,并存在胚胎败育现象,正常胚胎只占25.8%,导致四倍体结实率比二倍体降低22.6%.     

光敏核不育水稻短日照植株的雄性器官和育性异常

采用形态学方法比较观察了 10个水稻 (OryzasatavaL .)雄性核不育系的雄性器官的发育状况 :1)雄性核不育系短日照植株的自交结实率和花粉可育度均显著低于对照 ;2 )程度不同地存在花药开裂和散粉障碍 ,有多寡不等的柱头无任何花粉附着 ,且附着在不育系柱头上的花粉萌发力弱 ;3)胚囊发育状况及异交结实率均未见异常。说明光敏核不育水稻短日照植株自交结实率低下乃因雄性核不育基因表达所致。     

雄性不育无籽刺梨花药发育过程中生理特性的初步研究

为探究无籽刺梨(Rosasterilis)雄性不育原因,采用1%I2-KI染色法观察花粉活性,并对无籽刺梨和正常可育刺梨(R.roburghii)花药不同发育时期的生理生化指标进行了研究。结果表明,无籽刺梨的败育花粉占95.5%,刺梨的正常花粉占99%。刺梨花药的可溶性蛋白质、可溶性糖和脯氨酸含量在各时期的总体变化趋势相似,可溶性淀粉含量呈上升趋势,而无籽刺梨花药的可溶性蛋白质、可溶性糖、淀粉和脯氨酸含量在各时期的变化无规律可循,且花粉成熟期这4种物质的含量均明显低于刺梨,即花粉成熟期缺少各营养物质的积累。在花药发育过程中,无籽刺梨的SOD活性均低于刺梨;MDA含量呈上升趋势,且上升幅度比刺梨大;MDA含量和POD活性均高于刺梨。因此,营养物质的匮乏和酶系统的紊乱可能是导致无籽刺梨雄性不育的原因。     

矮慈姑的传粉机制与交配系统

对矮慈姑（ＳａｇｉｔａｒｉａｐｙｇｍａｅａＭｉｑ）的传粉过程与花粉流作了观察,用同工酶遗传标记法对其一个自然居群的异交率作出定量估计。在自然及人工居群中均观察到虫媒传粉,其中有较大比例的近距离传粉。未检出风媒花粉流。自交可育,同时存在雌雄同株和花序内雌雄花异熟等异交机制。异交率估计结果（ｔ＝４９９％）表明其交配系统为异交／自交兼性系统。作者认为,其较大比例的自交应归因于同株之不同克隆分株间交配的频繁发生     

琯溪蜜柚和度尾蜜柚自交和互交胚珠发育状况的比较

用石蜡切片技术对溪蜜柚 (Citrus grandis cv.Guanxim iyou)和度尾蜜柚 (C.grandis cv.Duweimiyou)自交和互交胚珠发育进行了比较观察。溪蜜柚的自交授粉后胚珠发育有 2种不同情形 (I型和 II型 ) , 型在授粉后5 d的珠心组织细胞有变形现象 ,接着胚囊萎缩 ,但外珠被能生长发育到 5 0 d左右 ; 型受精后胚乳发育异常 ,种子不能发育。度尾蜜柚自交胚囊中胚乳没有出现 ,在授粉后 35 d左右珠心组织和囊腔同时萎缩。互交胚珠发育正常 ,成熟果实有籽。表明两品种是自交不亲和的 ,导致了果实无籽。但两者的识别反应和阻抑部位不同 ,溪蜜柚 I型可能出现在受精前 , 型出现在受精后 ,而度尾蜜柚自交花粉管可能没有进入胚囊 ,是一种受精前的障碍。瘪籽的大小与不亲和障碍发生的时期有关。实验为进一步研究两品种的自交不亲和性遗传机理提供了实验证据 ,将有助于解决由于果实无籽导致的汁胞粒化、裂果、裂瓣等品质问题。     

Guan溪蜜柚和度尾蜜柚自交和互交胚珠发育状况的比较

用石蜡切片技术对Guan溪蜜柚（Citrus grandis cv.Guanximiyou)和度尾蜜柚（C.grandiscv.Duweimiyou)自交和互交胚珠发育进行了比较观察。Guan蜜柚的自交授粉后胚珠发育有2种不同情形（Ⅰ型和Ⅱ型）,I型在授粉后5d的珠心组织细胞有变形现象。接着胚囊萎缩,但外珠被能生长发育到50d左右;Ⅱ型受精后胚乳发育异常,种子不能发育,度尾蜜柚自交胚囊中胚乳没有出现,在授粉后35d左右珠心组织和囊腔同时萎缩,互交胚珠发育正常。成熟果实有籽。表明两品种是自交不亲和的,导致了果实无籽。但两者的识别反应和阻抑部位不同,Guan溪蜜柚I型可能出现在受精前,Ⅱ型出现在受精后,而度尾蜜柚自交花粉管可能没有进入胚囊,是一种受精前的障碍。瘪籽的大小与不亲和障碍发生的时期有关,实验为进一步研究两品种的自交不亲和性遗传机理提供了实验证据。将有助于解决由于果实无籽导致的汁胞粒化,裂果,裂瓣等品质问题。     

小麦×大麦胚胎发育的研究——Ⅲ.小麦×大麦胎胚发育过程中淀粉的积累和动态

我们观察了小麦与大麦杂交胚胎发育过程中,雌蕊各个组成部分发生的淀粉积累和转移的动态。结果如下: 1.胚乳和杂种胚早期发育过程中,子房壁和胚囊中淀粉的积累和动态的趋势与其他学者所作小麦自交的情况基本相同。2.当胚乳细胞充满胚囊而胚没有分化时,子房壁中的淀粉已极少,当胚乳解体时,子房壁中淀粉已几乎消失。可见,子房壁中淀粉的迅速消失与胚乳的迅速败育是平行的。3.胚囊发育的停滞与败育跟子房壁组织中淀粉的积累及对胚囊营养的正常供应有密切关系。4.花柱、珠被、珠心组织及胚囊中的助细胞和反足细胞,在整个杂种胚和胚乳发育过程中,始终不存在淀粉粒。助细胞胚亦和助细胞一样,无淀粉粒的存在。     

小麦×大麦胚胎发育的研究——Ⅲ.小麦×大麦胎胚发育过程中淀粉的积累和动态

我们观察了小麦与大麦杂交胚胎发育过程中,雌蕊各个组成部分发生的淀粉积累和转移的动态。结果如下: 1.胚乳和杂种胚早期发育过程中,子房壁和胚囊中淀粉的积累和动态的趋势与其他学者所作小麦自交的情况基本相同。2.当胚乳细胞充满胚囊而胚没有分化时,子房壁中的淀粉已极少,当胚乳解体时,子房壁中淀粉已几乎消失。可见,子房壁中淀粉的迅速消失与胚乳的迅速败育是平行的。3.胚囊发育的停滞与败育跟子房壁组织中淀粉的积累及对胚囊营养的正常供应有密切关系。4.花柱、珠被、珠心组织及胚囊中的助细胞和反足细胞,在整个杂种胚和胚乳发育过程中,始终不存在淀粉粒。助细胞胚亦和助细胞一样,无淀粉粒的存在。     

无籽罗汉果选育的初步研究

采用多倍体诱导、不同倍性配子杂交和组织培养等技术方法,培育出三倍体无籽罗汉果新种质类型。通过与当前罗汉果主要品种的对比栽培实验和观测,表明无籽罗汉果优良株系花、叶等器官形体较大,植株长势健旺,现蕾开花较早,果实无籽或极少籽,甜苷Ⅴ含量比其母本提高36.28%。其生物学性状的变化,尤其是甜甙含量的提高和抗性的增强,表现出了重大的开发利用价值。就无籽罗汉果的生育周期、形态特征、生长发育规律以及果实品质进行初步报道。     

Embryogenesis and plant regeneration of hot pepper (<Emphasis Type="Italic">Capsicum annuum</Emphasis> L.) through isolated microspore culture

We report high frequencies of embryo production and plant regeneration through isolated microspore culture of hot pepper (Capsicum annuum L.). Microspores cultured in modified NLN medium (NLNS) divided and developed to embryos. Globular and heart-shaped embryos were observed from 3 weeks after the beginning of culture, and many embryos reached the cotyledonary stage after 4 weeks of culture. These cotyledonary embryos developed to plantlets after transfer to solid B5 basal medium. We also optimized conditions for embryo production by varying the pretreatment media, the carbon sources, and culture densities. Heat shock treatment in sucrose-starvation medium was more effective than in B5 medium. Direct comparisons of sucrose and maltose as carbon sources clearly demonstrated the superiority of sucrose compared to maltose, with the highest frequency of embryo production being obtained in 9% (w/v) sucrose. Microspore plating density was critical for efficient embryonic induction and development, with an optimal plating density of 8 × 10⁴–10 × 10⁴/ml. Under our optimized culture conditions, we obtained over 54 embryos, and an average of 5.5 cotyledonary embryos when 10 × 10⁴ microspores were grown on an individual plate.     

On the Embryonic Development and Ovule Culture of Interspecific Hybrids Between Populus simonii Carr and P.pyramidalis Borkh

The embryonic development following P. simonii Cart. × P. pyramidalis Borkh. is described in the present paper. The majority of pollen grains of P. pyramidalis Borkh. may germinate on the stigma of P. simonii Cam and the pollen tubes grow normally through the style and enter the embryo sac from the micropyle. Fertilization occurs as usual 4–7 days after pollination. A lot of proembryos and heart-shaped embryos are abortive; however, the others may develop normally and grow into mature embryos. Some of the endosperms appear normal and others may degenerate at free nuclear stage or cease to develop further at cellular stage. The ovules containg immature hybrid embryos of 19 days, 22 days, 26 days and 29 days after pollination at various developmental stages (heart-shaped stage, torpedo-stage and cotyledonary elongation stage) are excised and inoculated on nutrient agar for culture. The results show that: ( 1 ) Mll0 medium ( 1/2 MS+IAA 0.01 mg/L+BA 0.1 mg/L+sucrose 2% ) is the best of all the media used; (2) immature hybrid embryos of various developmental stages contained in ovules cultured in vitro may grow into normal plantlets.     

Isolation and identification of substances inducing formation of tracheary elements in cultured carrot-root slices

Data are presented on the cytokinin status of seeds and seed components, at different stages of development in Phaseolus coccineus L., as determined with the soybean callus growth bioassay: A change in cytokinin types according to developmental stage occurred: from biologically very active less polar types (zeatin=Z) at early stages to more polar types (zeatin glucoside=Z₉G and zeatin riboside=Zr), with relatively low biological activity, at intermediate and late stages of seed development: When cytokinins were analyzed separately in embryos (embryo proper) and suspensors at two embryonic stages: heart-shaped (A) and middle cotyledonary embryos (stage B) respectively, it was found that: i) at stage A, the suspensor showed cytokinin activity at the level of Z, 2iPA (2-isopentenyladenosine) and Zr, whereas more polar cytokinins (Z₉G, Zr) were present in the embryo; ii) at stage B, when the embryo seems to become autonomous for cytokinin supply, there was a relative abundance of active cytokinins (Z, 2iPA) in the embryo to which Z₉G activity in the suspensor corresponded. It is concluded that the suspensor plays an essential role in embryogenesis by acting as a hormone source to the early embryo.Abbreviations GA gibberellic acid - 2iPA 2-isopentenyladenosine - Stage A heart-shaped embryo - siage B middle cotyledonary embryo - Z zeatin - Z₉G zeatin glucoside - Zr Zeatin riboside     

Anatomical study of zygotic and somatic embryos of Tilia cordata

A comparative anatomical study was carried out on zygotic and somatic embryos of Tilia cordata Mill. to evaluate the effect of growth conditions on their development. Zygotic embryos (heart-shaped, torpedo, cotyledonary), collected during two autumn periods, were examined to investigate the effect of growing season on embryo development. In comparison, the influence of growth conditions on the development of somatic embryos in vitro was also studied. Treatment with abscisic acid (ABA) and polyethylene glycol-4000 induced the development of somatic cotyledonary embryos similar to zygotic embryos with respect to morphology and anatomy, as illustrated by the differentiation of the apical meristems and of the procambium. The pattern of accumulation of starch and protein was also similar in these embryos. Somatic cotyledonary embryos that developed spontaneously without ABA showed defective accumulation of storage material and a general failure to form the shoot apical meristem, leading to very low germination rates. Vacuolar phenolic deposits were observed along the procambium of both zygotic and somatic embryos regardless of the maturation stage. Tracheid formation was observed only in somatic embryos formed without ABA in the medium and in precociously germinated somatic embryos. Phenolic vacuolar inclusions were frequently observed in epidermal cells of these embryos. This revised version was published online in June 2006 with corrections to the Cover Date.     

Metabolic regulation in C4 photosynthesis: Phosphoenol pyruvate carboxylase and 3C intermediates of the photosynthetic carbon reduction cycle

Summary Gibberellins and auxins were extracted from embryos and suspensors of Phaseolus coccineus L. at two stages of development: A) heart-shaped embryo and B) cotyledonary embryo with suspensor in the initial stage of degeneration. The time interval between the two stages was 5–6 days.In both embryos and suspensors, gibberellin (GA)-like activity was found in three fractions: F-1 (ethyl acetate fraction at pH 8.0), F-2 (free GAs) and F-3 (bound GAs). At stage A, the total GA activity in the suspensor was about 30 times greater than in the embryo and the bound GAs contributed by about 90% to the total GA content. A dramatic decrease in level of bound GA-like substances was found in suspensors at stage B, when the level of total GAs in the embryo had increased to 10 times that at stage A. This might suggest a transport of GAs from the suspensor to the embryo. In both embryo and suspensor, qualitative changes in GAs with shift in activity of the fractions tested occurred at the two developmental stages.The methanolic extracts of stage A suspensors showed two inhibitors, one much more active than the other, and two large peaks of growth promoting activity at Rf 0.4–0.7; in stage A embryos, the general activity of the extracts was lower and the promoting effect was spread over Rf 0.3–0.9.The present results seem to support the view that the suspensor plays a role in embryogenesis by acting as a site of synthesis of growth regulators needed by the embryo.Abbreviations F-1 ethyl acetate fraction at pH 8.0 - F-2 free gibberellins - F-3 bound gibberellins - GA gibberellic acid - Stage A heart-shaped embryo - stage B cotyledonary embryo with suspensor in the initial stage of degeneration     

杂交柑桔的胚胎发育(英文)

柑桔类珠心胚现象对研究其进化和种质保存方面有着重要意义,但是给其有性杂交育种工作造成相当大的麻烦：它不仅使杂交后代难于区分合子苗和珠心苗,而且会使合子胚发育中途天折。解决的途径是适时分离合子胚进行单独培养。为此,本工作于柑桔胚胎发育的不同时期取样、固定、观察,研究其合子胚发育规律,确定珠心胚侵入胚囊的时期。结果表明：一般情况下,授粉后３０ｄ,合于处于单细胞状态（Ｆｉｇ．１Ａ）;授粉后４０ｄ;合子开始分裂（Ｆｉｇ．１Ｂ）;授粉后４５ｄ,合子胚形成８－１６个细胞,珠心胚开始分裂（Ｆｉｇ．１Ｃ）;授粉后５０ｄ,合子形成球形胚,珠心胚尚未侵入胚囊（Ｆｉｇ．１Ｄ＆Ｅ）;授粉后５５ｄ,合子胚为球形胚或早心形胚,开始有少数珠心胚侵入胚囊（Ｆｉｇ．１Ｄ＆Ｅ）;授粉后６０ｄ,合子胚为心形胚（Ｆｉｇ．１Ｇ）。授粉后８０ｄ,合子胚为晚心形,珠心胚巨大量侵入并迅速发育（Ｆｉｇ．１Ｈ）。杂交的组合对合子,胚珠及果实发育均有一定影响,主要取决于杂交亲本。与种间杂交相比,属间杂交其合子分裂较迟,合子胚发育较慢,珠心胚侵入时期也较迟（Ｔａｂｌｅｌ,Ｆｉｇ．２）。这种发育速度的差异,引起所产生的合子胚的大小（Ｔａｂｌｅ２）珠心胚侵率（Ｔａｂ     

Turnover of cell-wall polysaccharides during somatic embryogenesis and development of celery (Apium graveolens L.)

Non-embryogenic cells (NEC) and embryogenc cells (EC) were separated from cell clusters derived from the hypocotyl segments of celery seedlings, which had been suspension-cultured in MS medium supplemented with 10⁵ M 2,4-D. The EC formed globular embryos in medium without 2,4-D. The globular embryo developed through heart-shaped, torpedo to cotyledonary embryos within 10 days. The EC and developing embryos were fractionated into symplastic [MeOH, hot water (HW), starch (S)] and apoplastic [pectin, hemicellulose, TFA (trifluoroacetic acid)-soluble and cellulose] fractions. The EC contained lower levels of sugar in the MeOH fraction and higher levels of starch than NEC. In the apoplastic fractions, there were no differences of total sugar amounts between NEC and EC. Cellulose contents were about 10% of the wall polysaccharides. During somatic embryogenesis, total sugar contents of the MeOH and HW fractions increased till the heart-shaped embryo stage, and then decreased during the torpedo and cotyledonary embryo stages. The sugar contents of the starch, pectin, TFA-soluble, and cellulose fractions did not change during the stages mentioned above. However, the hemicellulose substances remarkably increased during embryogenesis, and then decreased as the development proceeded. The neutral sugar components of the hemicellulosic fractions were analyzed. Arabinose increased markedly in EC to the globular embryo stage, but decreased as the development proceeded. Galactose increased only at the torpedo and cotyledonary embryo stages. Xylose was present at lower levels in all stages of embryogenesis than in the differentiated hypocotyl cell walls. These results suggest that there was a high turnover of arabinogalactan polysaccharides during embryogenesis, and that xylan accumulated in the cell walls of differentiated cells     

Role of enzymes and identification of stage-specific proteins in developing somatic embryos of chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L.)

Accumulation of proline, activities of peroxidase (POX), catalase (CAT), phenylalanine ammonia lyase (PAL) and malate dehydrogenase (MDH) were studied during different developmental stages of somatic embryos in chickpea. Callus cultures that did not form somatic embryos served as control. While increased levels of proline and POX activity were noticed in globular stages of embryos, CAT activity increased during early and late heart-shaped embryo formation indicating tissue-specific activation of these enzymes. The activity of PAL reached a peak during torpedo and cotyledonary stages of embryo development. On the other hand, MDH activity enhanced during the germination of somatic embryos inferring more requirement of energy during this stage. Electrophoretic (sodium dodecyl sulfate polyacrylamide gel electrophoresis) pattern of proteins revealed that ten bands are associated with non-embryogenic tissues, whereas 11 bands with globular, heart, torpedo and cotyledonary stages of embryo development and nine bands during the germination stage of embryos. Two extra stage-specific protein bands with molecular masses of 16 and 18 kDa appeared during globular, heart, torpedo, and cotyledonary stages. But, these bands disappeared during germination of embryos and are absent in non-embryogenic cultures. This study thus may help in the identification of proteins and the role of above enzymes during different developmental stages of somatic embryo induction and their maturation in a recalcitrant leguminous crop plant chickpea.     

黄顶菊的大孢子发生、雌配子体与胚胎发育

用常规石蜡制片对黄顶菊(Flaveria bidentis(L.) Kuntze)大孢子发生、雌配子体和胚胎的发育过程进行了观察.黄顶菊雌蕊柱头二裂,2心皮,1室,单胚珠,基生胎座,单珠被,薄珠心,倒生胚珠,具发达的珠被绒毡层.珠心表皮下分化出孢原细胞,孢原细胞直接发育为大孢子母细胞,大孢子母细胞减数分裂形成直列四分体...     

多胚水稻品系APⅣ不同类型胶囊的受精及其胚胎形成

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