植物无融合生殖研究进展

本文综述了植物无融合生殖研究进展。无融合生殖能固定杂种优势,是新的研究热点。无融合合生殖转充研究取得长足进展。胚胎发生研究手段由切片技术逐渐发展为整体透明、组化荧光技术。大孢子母细胞（ＭＭＣ）细胞壁无胼胝质（ｃａｌｌｏｓｅ）及ＭＭＣ哑铃状核是二倍性孢子形成区别于有性生殖的特征。ＤＮＡ分子标记是无融合生殖研究的新的有效工具,狼尾草属、摩擦禾属的无融合生殖分子标记已被找到,并且后者已定位到玉米第６染色     

高梁SSA—1无融合生殖胚胎学研究

经常规石蜡切片法,在光学水平观察了高染（Ｓｏｒｇｈｕｍｂｉｃｏｌｏｒｂｉｃｏｌｏｒ．）Ｍｏｅｎｃｈ）ＳＳＡ－１无融合生殖的胚胎发生。高染ＳＳＡ－１的无融合生殖为无孢子生殖和二倍体孢子生殖两种类型。两种生殖类型的单核胚囊经３次有丝分裂成７细胞（８核）的成熟胚囊,由卵细胞、２个助细胞、２个极核和３个足细胞组成,反足细胞迅速分裂增殖,形成由２０－３０个细胞组成的细胞团。此外,还具有一定频率的无孢子生殖多     

禾本科植物无融合生殖(综述)

禾本科植物包含了世界上最重要的农作物,也包含了最多的无融合生殖的种类,通过无融合生殖可将农作物的F1代杂种优势固定下来,这在固定农作物杂种优势的利用上具有巨大的潜力,然而禾本科植物无融合生殖作为其繁殖多样性的一种形式,在系统进化过程中的作用是非常复杂的,本文统计了禾本科无融合生殖的分布,概述了其无融合生殖的细胞学,遗传学和分子生物学研究进展。     

非洲狼尾草无融合生殖胚胎学研究

报道非洲狼尾草（ＰｅｎｎｉｓｅｔｕｍｓｑｕａｍｕｌａｔｕｍＦｒｅｓｅｎ）的胚囊形成、胚胎发生与发育过程。非洲狼尾草的孢原细胞直接发育成大孢子母细胞,并由它分裂产生三分体。从大孢子母细胞发育至三分体的不同阶段,均会出现败育。性细胞退化期间,其周围的珠心组织中,常出现一至多个体积较大的无孢子生殖原始细胞。通常只有靠近珠孔端的１个无孢子生殖原始细胞体积进一步增大,并出现大液泡,发育成无孢子生殖单核胚囊。随后,其核经连续两次有丝分裂,形成无孢子生殖四核胚囊,胚囊内的４个核常聚积在珠孔端,４个核进一步分化形成１个卵细胞、１个助细胞和具两个极核的中央细胞,没有反足细胞。胚囊发育属于大黍型。其它的无孢子生殖原始细胞能发育到单核或二核胚囊阶段,而后核解体导致胚囊败育。胚的发生有两种类型：（１）早发生胚。大多数胚囊在开花前一、二天,次生核未分裂,卵细胞不经受精,自发分裂形成胚。（２）迟发生胚。少数胚囊的卵细胞不经过受精,但需要在开花后三、四天次生核分裂为多个胚乳核时才开始分裂。无论是早发生胚或迟发生胚,卵细胞在分裂前具有极性,珠孔端有大液泡,细胞质稀薄,合点端细胞质较浓。胚的发育经历球形胚、梨形胚和胚分化阶段。     

龙须草无融合生殖的胚胎学证据

采用石蜡切片技术对龙须草(Eulaliopsisbinata(Rotz)C.E.Hubb)进行了系统的胚胎学研究,证明龙须草为禾本科植物中一种新的无融合生殖材料。龙须草无融合生殖方式为无孢子生殖,在胚珠发育早期,多个珠心细胞特化为无孢子生殖原始细胞,由原始细胞发育为单核胚囊,经两次有丝分裂形成4核胚囊,进一步分化形成两种类型的成熟胚囊:(1)具1个卵细胞,1个助细胞和2个极核,占观察总数的67.6%;(2)具1个卵细胞,2个助细胞和1个极核,占观察总数的32.4%。胚囊发育属大黍型。多个无孢子生殖原始细胞可以同时发育,最后形成2个或多个胚囊,其比例为17.7%。胚珠内没有有性胚囊的发育。胚的发生有两种类型:(1)早发生胚(74%),开花前1~2d,极核未分裂前卵细胞分裂形成胚;(2)迟发生胚(26%),开花后2~3d,极核分裂形成多个胚乳游离核后,卵细胞启动分裂形成胚。存在多胚现象,多胚来自不同胚囊内卵细胞的孤雌生殖,多胚发生率为13%。胚乳由极核不经受精自发分裂产生。     

龙须草无融合生殖的胚胎学证据

采用石蜡切片技术对龙须草(Eulaliopsis binata(Rotz)C.E.Hubb)进行了系统的胚胎学研究,证明龙须草为禾本科植物中一种新的无融合生殖材料.龙须草无融合生殖方式为无孢子生殖,在胚珠发育早期,多个珠心细胞特化为无孢子生殖原始细胞,由原始细胞发育为单核胚囊,经两次有丝分裂形成4核胚囊,进一步分化形成两种类型的成熟胚囊:(1)具1个卵细胞,1个助细胞和2个极核,占观察总数的67.6%;(2)具1个卵细胞,2个助细胞和1个极核,占观察总数的32.4%.胚囊发育属大黍型.多个无孢子生殖原始细胞可以同时发育,最后形成2个或多个胚囊,其比例为17.7%.胚珠内没有有性胚囊的发育.胚的发生有两种类型:(1)早发生胚(74%),开花前1～2 d,极核未分裂前卵细胞分裂形成胚;(2)迟发生胚(26%),开花后2～3 d,极核分裂形成多个胚乳游离核后,卵细胞启动分裂形成胚.存在多胚现象,多胚来自不同胚囊内卵细胞的孤雌生殖,多胚发生率为13%.胚乳由极核不经受精自发分裂产生.     

小麦和无融合生殖披碱草杂交后代（BC2F2）的无融合生殖及胚胎发育过程中的异常现象研究

对小麦（Triticum aestivum）和无融合生殖披碱草（Elymus rectisetus）的染 色体数目为42的杂种后代（BC2F2）单株进行了RAPD检测和胚胎学研究。RAPD检测结果表明：染色体数目为42条的BC2F2单株的遗传组成与普通小麦的遗传组成十分接近,但是在部分单株中出现了披碱草的特异带。由此可以推测,经过回交和自交后小麦草的部分染色体片段已经整合进了小麦的染色体。在部分BC2F2单株胚胎学切片中发现了较高比例的（5%左右）双孢原、早发胚以及多胚囊等无融合生殖现象,直接表明了无融合生殖基因转移。由于基因整合的多样性,无融合生殖基因在有些单株中并没有充分表达,从而造成了某些单株胚胎发育的异常。     

无融合生殖

自1745年博内(Bonnet)在研究蚜虫中发现孤雌生殖以来,世界各地的植物学家,也在众多的植物中发现了这种奇特的生殖现象,迄今为止,已记录到36科300余种植株的孤雌生殖。为同有性生殖和无性生殖相区分,植物学家把它定名为无融合生殖。 由于无融合生殖生理机制十分复杂,长期以来,对其概念的界定,植物学界一直没有统一。我国学者蔡得田和陈冬玲,在1993年提出:无融合生殖是指发生在植物胚珠中,不经过精卵融合形成胚,以种子进行繁殖的生殖方式。此界定包括三部分内容,一是它与无性生殖明显的区别是发生在性器官中;二是没有性融合过程而区别于有性生殖;三是以种子繁殖     

水蔗草兼性无融合生殖胚胎学研究

对水蔗草 (ApludamuticaL .)的生殖方式进行研究 ,结果表明水蔗草进行兼性无融合生殖。胚囊发育分为两种类型 ,即有性生殖的蓼型和无孢子生殖的大黍型。无融合生殖胚囊频率为 6 0 .74%。在大孢子母细胞发育至四分体后 ,珠孔端的 3个大孢子解体。合点端的大孢子未解体时 ,邻近大孢子的 1个珠心细胞开始特化 ,形成无融合生殖的原始细胞 ,由该原始细胞发育形成有 1个卵细胞、1个助细胞和 2个极核的四核胚囊。     

水蔗草兼性无融合生殖胚胎学研究

对水蔗草(Apluda mutica L.)的生殖方式进行研究,结果表明水蔗草进行兼性无融合生殖.胚囊发育分为两种类型,即有性生殖的蓼型和无孢子生殖的大黍型.无融合生殖胚囊频率为60.74%.在大孢子母细胞发育至四分体后,珠孔端的3个大孢子解体.合点端的大孢子未解体时,邻近大孢子的1个珠心细胞开始特化,形成无融合生殖的原始细胞,由该原始细胞发育形成有1个卵细胞、1个助细胞和2个极核的四核胚囊.     

大黍(Panicum maximum Jacq.)的胚珠附器

观察了大黍(Panicum,maximum Jacq．)胚珠附器的发生时间、位置和发育过程及其细胞化学特征。结果显示：(1)大孢子母细胞时期,珠孔端有一个或多个珠心表皮细胞开始伸长、膨大,特化为胚珠附器。(2)当胚珠附器伸长、膨大至最大程度时,胚珠附器细胞表现出显著的极性特征：细胞核位于细胞的珠孔端,大而清晰;细胞内同时形成了一个特大的液泡,几乎占据了整个细胞的合点端;细胞质则被挤到珠孔端一侧,集中分布在核的周围。(3)胚珠附器从开始出现到发育成熟,都没有淀粉粒的积累;但是,PAS反应显示胚珠附器细胞壁和细胞质都比普通珠心细胞的染色程度深,这说明其细胞壁和细胞内部富含可溶性多糖。     

外来植物坚尼草生态学研究

坚尼草(Panicum maximum Jacq.)是一种外来草本植物,已逸为野生,扩展迅速,对本地草本植物构成一定的影响。其扩展能力与其生长和繁殖特点有很大关系。研究表明,坚尼草对环境的适应能力较强;植株较高大,生长迅速,有较强的竞争能力;花期长,单丛坚尼草从始穗到终穗,花期可长达6～7个月;同一穗的果实熟期不同,而且边熟边脱落,从抽穗的5～7d起开始有果实脱落,脱落时间可持续1个月左右,以抽穗2～3周为脱落高峰;不同时间成熟的果实,种子萌发率也不同,7月份成熟的果实比9月份成熟的果实萌发率高。同一穗中,以抽穗后2～3周（果实脱落的高峰期）成熟的果实萌发率较高。果实成熟后,不能立即萌发,储存1～6个月,种子萌发率较高。     

外来植物坚尼草生态学研究

坚尼草（Panicm maximum Jacq.^-）是一种外来草本植物,已逸为野生,扩展迅速,对本地草本植物构成一定的影响。其扩展能力与其生长和繁殖特点有很大关系。研究表明,坚尼草对环境的适应能力较强;植株较高大,生长迅速,有较强的竞争能力;花期长,单丛坚尼草从始穗到终穗,花期可长达6－7个月;同一穗的果实熟期不同,而且边熟边脱落,从抽穗的5－7d起开始有果实脱落,脱落时间可持续1个月左右,以抽穗2－3周为脱落高峰;不同时间成熟的果实,明发率也不同,7月份成熟的果实比9成熟的果实萌发率高。同一穗中,以抽穗后2－3周（果实脱落的高峰期）成熟的果实萌发率较高。果实成熟后,不能立即萌发,储存1－6个月,种子萌发率较高。     

Chromosomal and embryological analyses in sexual x apomictic hybrids ofPanicum maximum Jacq.

Summary Cytological analyses in series of crosses between 7 sexual pistillate and 8 apomictic staminate parents of speciesPanicum maximum (Gramineae) are reported. Although these 15 progenitors were tetraploid (2n = 32), 2 dihaploids (2n = 16), 45 hexaploids (2n = 48) and 5 octoploids (2n = 64) were observed among 333 progeny plants. The role of unreduced gametes as the originators of polyploidy is discussed in relation to the so-called elements of apomixis. The 2 dihaploids appeared to be sexual while the hexaploids and octoploids were all apomictic. At the tetraploid level sexual and apomictic hybrids segregated in a ratio close to 11. These results were then compared to those already obtained from studies on other tropical grasses and indicate a simple genetic determinism for gametophytic apomixis.     

Somatic embryogenesis and plant regeneration from leaf tissues of Panicum maximum Jacq.

Summary Somatic embryogenesis was induced in proliferating leaf segments of Panicum maximum Jacq., cultured on Murashige and Skoog's nutrient medium containing 2,4-dichlorophenoxyacetic acid and coconut milk. The embryoids gave rise to plants on a medium containing gibberellic acid. The plants were successfully transplanted to soil and grown to maturity. Histological examination of proliferating leaves showed that the embryogenic callus tissue was formed by divisions in cells of the lower epidermis as well as the mesophyll tissue. The regenerated plants showed the normal chromosome number of 2n=4x=32.Florida Agriculture Experiment Station Journal Series No. 2775     

Isozyme polymorphism and organization of the agamic complex of the Maximae (Panicum maximum Jacq., P. infestum Anders,and P. trichocladum K. Schum.) in Tanzania

The tribe Maximae (Panicum maximum Jacq., P. infestum Anders., P. trichocladum K. Schum.) includes two sympatric pools with different modes of reproduction and ploidy levels: an apomictic and tetraploid pool on the one hand, and a smaller, sexual and diploid pool on the other. From an analysis of isozyme polymorphism five main results were evident. First, overall polymorphism is considerable showing that apomixis does not lead to a reduction in diversity. Second, the isozyme polymorphism of the two pools is similar, and this may be explained by reciprocal gene flow (low, but continuous) between these two pools. Third, maximum local polymorphism is due to the simultaneous presence of P. maximum sexuals and P. infestum apomicts. A continuum exists between the two species. Fourth, a high proportion of rare alleles, arising from introgression from P. infestum, characterized the isozyme polymorphism. These rare alleles, strongly counter-selected at the diploid level, are maintained by apomixis; the frequency of triplex or quadruplex genotypes was nevertheless low. Fifth, the heterozygosity level within apomicts is not higher than that of sexuals, showing that the apomixis-polyploidy combination does not lead to a higher frequency of very heterozygous individuals.     

Molecular Diversity and Genetic Structure of Guineagrass (Panicum maximum Jacq.), a Tropical Pasture Grass

Guineagrass (Panicum maximum Jacq.) is a forage grass found in tropical and subtropical regions. It is an apomictic and tetraploid species from Africa. The objective of this study was to evaluate the genetic diversity of guineagrass accessions sampled from its regions of origin, which is in Tanzania and Kenya. In this study, a total of 396 accessions were analyzed, and a collection of reproducible and informative microsatellites was developed. Thirty microsatellites were employed to characterize these accessions. A total of 576 clones were sequenced from microsatellite-enriched libraries. Flanking primers were designed for 116 microsatellite loci and screened using a sample of 25 guineagrass accessions. The thirty selected polymorphic microsatellites employed in this study produced a total of 192 bands when evaluated in the 396 P. maximum accessions, with an average of 6.4 bands per microsatellite. Four genetic clusters were identified in the collection using STRUCTURE analysis, and these results were confirmed using AMOVA. The largest genetic variation was found within clusters (65.38%). This study revealed that the collection of accessions from the P. maximum region of origin was a rich source of genetic variability. The geographical distances and genetic similarities among accessions did not indicate a significant association between genetic and geographical variation, supporting the natural interspecific crossing between P. maximum, P. infestum and P. trichocladum as the origin of the high genetic variability and the existence of an agamic complex formed by these three species.     

The effect of nitrogen and phosphorus on starch accumulation and net photosynthesis in two variants of Panicum maximum Jacq.

Abstract. Two anatomical variants of Panicum maximum Jacq. were observed to accumulate an unusually large number of starch grains in the bundle sheath chloroplasts when grown under controlled environmental conditions in a nutrient medium containing a low level of nitrate nitrogen (20 mg N dm⁻³ as KNO₃). When these plants were placed under dark conditions the chloroplasts were destarched, but exhibited a marked distortion of the thylakoid membranes. Under a higher level of nitrate nitrogen supply (200 mg N dm⁻³ as KNO₃) the number of starch grains was markedly reduced compared to that observed above in both plant variants. When the nitrogen was supplied as ammonium nitrogen (200 mg N dm⁻³ as NH₄Cl) there was again a high level of starch in the bundle sheath chloroplasts, the level being only slightly lower than that observed at the low KNO₃ supply. An unusually large number of starch grains accumulated in the bundle sheath chloroplasts in the absence of added phosphorus in the nutrient medium, in the presence of the higher nitrate nitrogen level. It is suggested that the increased starch accumulation results from a reduced trans-location of Calvin cycle intermediates out of the chloroplasts into the cytoplasm and that both nitrate nitrogen and phosphorus may play an important role in this process. A good correlation between high net photosynthetic activity and low bundle sheath starch content was observed. Nutrient medium requirements favouring low starch content in chloroplasts also favoured high net photosynthetic rates.     

Embryological Studies on Apomixis in Pennisetum squamulatum

Studies on the formation and development of the embryo sac of the apomictic material of Pennisetum squamulatum Fresen indicated that normal archesporial cell did form with consequent development of a megaspore mother cell and later meiotic division to give rise to a triad. But invariably the megaspore mother cell and the triad underwent degeneration after formation. During the period of formation or degeneration of the megaspore or the triad a number of nucellar cells around the degenerated sexual cell became much enlarged. Frequently, one of the enlarging nucellar cells near the micropylar end became vacuolated and then developed into an aposporous uninucleate embryo sac, which underwent two further mitotic divisions to form an aposporous four-nucleate embryo sac, where the four nuclei remained in the micropylar end. Thus in the mature aposporous embryo sac there were one egg cell, one synergid and one central cell (containing two polar nuclei). Antipodal cells were completely lacking. The pattern of development of the aposporous embryo sac resembles the panicum type. There were two types of embryo formed during apomictic development namely ( 1 ) The pre-genesis embryo--embryo formed without fertilization, 1 to 2 days before anthesis, and (2) The late-genesis embryo--derived from the unfertilized egg cells, 3 to 4 days after anthesis. In the late-genesis embryo type, the egg cell divided after the secondary nucleus has undergone division to form the endosperm nuclei. All egg cells developed vacuoles before they differentiated into embryos. The development of the aposporous embryo followed the sequence of the formation of globular, pearshaped embryo and full stages of differentiation. The unfertilized secondary nucleus divides to form free endosperm nuclei after being stimulated by pollination. The development of the endosperm belongs to the nuclear-type.     

雀稗属无融合生殖研究进展

雀稗属(Paspalum)为禾本科黍亚科多年生或一年生植物,是黍亚科内最有经济价值的类群之一。雀稗属植物种群极其复杂,大多数为多倍体。由于多倍体的存在及有性生殖的自交不亲和等原因,雀稗属植物表现出复杂多样的生殖特性,是禾本科中具备无融合生殖特性种类最多的属。对雀稗属无融合生殖的分布、无融合生殖相关的细胞学和胚胎学基础、无融合生殖的特点及其遗传学和分子生物学研究进展进行了综述。