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Asymmetric cell division of rice zygotes located in embryo sac and produced by in vitro fertilization. 位于胚囊内的水稻离体受精受精卵的不对称细胞分裂。
Pub Date : 2010-09-01 Epub Date: 2010-01-07 DOI: 10.1007/s00497-009-0129-9
Akiko Sato, Kiminori Toyooka, Takashi Okamoto

In angiosperms, a zygote generally divides into an asymmetric two-celled embryo consisting of an apical and a basal cell. This unequal division of the zygote is a putative first step for formation of the apical-basal axis of plants and is a fundamental feature of early embryogenesis and morphogenesis in angiosperms. Because fertilization and subsequent embryogenesis occur in embryo sacs, which are deeply embedded in ovular tissue, in vitro fertilization of isolated gametes is a powerful system to dissect mechanisms of fertilization and post-fertilization events. Rice is an emerging molecular and experimental model plant, however, profile of the first zygotic division within embryo sac and thus origin of apical-basal embryo polarity has not been closely investigated. Therefore, in the present study, the division pattern of rice zygote in planta was first determined accurately by observations employing serial sections of the egg apparatus, zygotes and two-celled embryos in the embryo sac. The rice zygote divides asymmetrically into a two-celled embryo consisting of a statistically significantly smaller apical cell with dense cytoplasm and a larger vacuolated basal cell. Moreover, detailed observations of division profiles of zygotes prepared by in vitro fertilization indicate that the zygote also divides into an asymmetric two-celled embryo as in planta. Such observations suggest that in vitro-produced rice zygotes and two-celled embryos may be useful as experimental models for further investigations into the mechanism and control of asymmetric division of plant zygotes.

在被子植物中,合子通常分裂成由顶细胞和基细胞组成的不对称的两细胞胚胎。合子的不平等分裂被认为是植物顶基轴形成的第一步,也是被子植物早期胚胎发生和形态发生的基本特征。由于受精和随后的胚胎发生在胚胎囊中,胚胎囊深埋在卵泡组织中,因此离体配子的体外受精是研究受精和受精后事件机制的有力系统。水稻是一种新兴的分子和实验模式植物,然而,对胚囊内第一次合子分裂的特征及其顶基胚极性的起源尚未深入研究。因此,在本研究中,首先通过对受精卵、受精卵和胚囊内的二细胞胚胎的连续切片观察,准确地确定了水稻合子在植物中的分裂模式。水稻合子不对称地分裂成两细胞胚,包括一个具有致密细胞质的较小的顶细胞和一个较大的液泡化的基底细胞。此外,对体外受精制备的受精卵的分裂谱的详细观察表明,受精卵也像植物一样分裂成不对称的双细胞胚胎。这些观察结果表明,体外产生的水稻受精卵和双细胞胚胎可能为进一步研究植物受精卵不对称分裂的机制和控制提供有用的实验模型。
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引用次数: 35
Genetic interaction between AINTEGUMENTA (ANT) and the ovule identity genes SEEDSTICK (STK), SHATTERPROOF1 (SHP1) and SHATTERPROOF2 (SHP2). AINTEGUMENTA (ANT)与胚珠鉴定基因SEEDSTICK (STK)、SHATTERPROOF1 (SHP1)和SHATTERPROOF2 (SHP2)的遗传互作
Pub Date : 2010-06-01 Epub Date: 2009-12-30 DOI: 10.1007/s00497-009-0130-3
Alessia Losa, Monica Colombo, Vittoria Brambilla, Lucia Colombo

AINTEGUMENTA (ANT) promotes initiation and growth of ovule integuments which cell fate is specified by ovule identity factors, such as SEEDSTICK (STK), SHATTERPROOF1 (SHP1) and SHATTERPROOF2 (SHP2). To study the genetic interaction between ANT and the ovule identity genes, we have obtained a stk shp1 shp2 ant quadruple mutant. The molecular and morphological characterization of the quadruple mutant and its comparison with the stk shp1 shp2 triple mutant, the shp1 shp2 ant triple mutant and the stk ant double mutant are here presented.

AINTEGUMENTA (ANT)促进胚珠被的形成和生长,而胚珠被的命运由胚珠身份因子如SEEDSTICK (STK)、SHATTERPROOF1 (SHP1)和SHATTERPROOF2 (SHP2)决定。为了研究ANT与胚珠同源基因之间的遗传相互作用,我们获得了一个株shp1 shp2的四重突变体。本文报道了该四重突变体的分子和形态特征,并与stk shp1 shp2三重突变体、shp1 shp2蚂蚁三重突变体和stk蚂蚁双重突变体进行了比较。
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引用次数: 21
Developmental and cytogenetic analyses of pollen sterility in Valeriana scandens L. 缬草花粉不育性的发育及细胞遗传学分析。
Pub Date : 2010-06-01 Epub Date: 2010-01-09 DOI: 10.1007/s00497-009-0128-x
Erica Duarte-Silva, André L L Vanzela, Jorge E A Mariath

Valeriana scandens presents perfect and pistillate flowers, the latter with sterile anthers. The species is composed of two varieties with different ploidy; V. scandens var. scandens (2n = 28) and V. scandens var. candolleana (2n = 56), both of which occur in RS, Brazil. Crosses between these varieties may give rise to hybrids with pollen sterility. In this study, we analyzed the microsporogenesis and microgametogenesis of sterile and fertile anthers, and also investigate whether pollen sterility is caused by an irregular meiotic process. Developmental analysis using light microscopy and scanning electron microscopy showed that sterile anthers develop similarly to fertile anthers until the end of meiosis. After this stage, sterile tetrads do not separate as a consequence of exine fusion between adjacent microspores, which is similar to sterile pollen of Brassica ms-cdl1 mutants. In addition, vacuolated immature pollen grains degenerate after separation. The cytogenetic analysis of the microspore mother cell (MMC) showed that the diploid population of V. scandens var. scandens (2n = 28) has pollen sterility that is not caused by a cytogenetic disturbance. The MMCs analyzed from prophase I to tetrad stage showed a regular meiotic process, indicating the phenotype of V. scandens sterile pollen is a postmeiotic process formed by fusion of exine between opposite microspores.

缬草呈现完美和雌蕊花,后者具有不育的花药。本种由两个倍性不同的变种组成;V. scandens var. scandens (2n = 28)和V. scandens var. candolleana (2n = 56),均发生在巴西RS。这些品种之间的杂交可能产生花粉不育的杂种。本研究分析了不育花药和可育花药的小孢子发生和小配子体发生,并探讨了花粉不育是否由不规则减数分裂过程引起。光镜和扫描电镜的发育分析表明,不育花药在减数分裂结束前与可育花药发育相似。在这一阶段之后,不育的四分体不会因为相邻小孢子之间的外胞融合而分离,这与芸苔属ms-cdl1突变体的不育花粉相似。此外,空泡化的未成熟花粉粒在分离后退化。对小孢子母细胞(MMC)的细胞遗传学分析表明,2n = 28的scandens var. scandens二倍体群体具有非细胞遗传学干扰引起的花粉不育性。从1前期到四分体的MMCs表现出规律的减数分裂过程,说明scandens不育花粉的表型是由对生小孢子之间的外壁融合形成的减数分裂后过程。
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引用次数: 9
Sperm dimorphism in terms of nuclear shape and microtubule accumulation in Cyrtanthus mackenii. 从核形状和微管积累的角度看金盏花精子二态性。
Pub Date : 2010-06-01 Epub Date: 2009-11-22 DOI: 10.1007/s00497-009-0123-2
Tomonari Hirano, Yoichiro Hoshino

Pollen tubes of Cyrtanthus mackenii, a species with bicellular pollen, were cultured in vitro to investigate nuclear phase changes during generative cell division and male germ unit (MGU) formation, using flow cytometric analysis. Results revealed that sperm cells were formed after 12 h of culture. During sperm maturation, the nuclei of sperm cells were not associated with the vegetative nucleus (unassociated sperm cells; Sua) and became longer than those of sperm cells associated with the vegetative nucleus (Svn). These findings indicate that the pair of sperm cells in the C. mackenii MGU is dimorphic in terms of nuclear shape. Dimorphism coincides with anti-alpha-tubulin antibody immunofluorescence, which was higher in the Sua than in Svn. Following treatment with oryzalin, triggering microtubule depolymerization, differences between nuclear shapes in the two sperm nuclei disappeared, suggesting that microtubule accumulation between sperm cells in the MGU correlates with differences in the nuclear shape.

采用体外培养双细胞花粉种Cyrtanthus mackenii花粉管,利用流式细胞术研究了生殖细胞分裂和雄性生殖单位(MGU)形成过程中的核相变化。结果表明,培养12 h后精子细胞形成。在精子成熟过程中,精子细胞核不与营养细胞核相关(非相关精子细胞;Sua),并且比与营养核相关的精子细胞(Svn)的长度更长。这些发现表明,在核形状方面,鲭鱼MGU的一对精子细胞是二态的。二态性与抗α -微管蛋白抗体免疫荧光一致,在Sua中高于Svn。在用米杂灵处理后,触发微管解聚,两个精子细胞核的核形状差异消失,提示MGU中精子细胞之间的微管积聚与核形状的差异有关。
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引用次数: 13
A comparative analysis of the distribution and composition of lipidic constituents and associated enzymes in pollen and stigma of sunflower. 向日葵花粉和柱头中脂质成分及相关酶的分布和组成比较分析。
Pub Date : 2010-06-01 Epub Date: 2009-12-15 DOI: 10.1007/s00497-009-0125-0
Rashmi Shakya, Satish Chander Bhatla

Spatial distribution and compositional analyses of the lipidic constituents in pollen and stigma of sunflower (Helianthus annuus L. cv. Morden) were conducted using ultrastructural, histochemical, and biochemical analysis. Detection of secretions at the base of stigmatic papillae and neutral lipid accumulations on the surface of stigmatic papillae and between adjacent pseudopapillae demonstrates the semidry nature of stigma surface in sunflower. Pollen coat is richer in lipids (8%) than stigma (2.2%) on fresh weight basis. Nile Red-fluorescing neutral lipids are preferentially localized in the pollen coat. Neutral esters and triacylglycerols (TAGs) are the major lipidic constituents in pollen grains and stigma, respectively. Lignoceric acid (24:0) and cis-11-eicosenoic acid (20:1) are specifically expressed only in the pollen coat. Similar long-chain fatty acids have earlier been demonstrated to play a significant role during the initial signaling mechanism leading to hydration of pollen grains on the stigma surface. Lipase (EC 3.1.1.3) activity is expressed both in pollen grains and stigma. Stigma exhibits a better expression of acyl-ester hydrolase (EC 3.1.1.1) activity than that of observed in both the pollen fractions. Expression of two acyl-ester hydrolases (41 and 38 kDa) has been found to be specific to pollen coat. Specific expression of lignoceric acid (24:0) in pollen coat and localization of lipase in pollen and stigma have been discussed to assign possible roles that they might play during pollen-stigma interaction.

向日葵花粉和柱头脂质成分的空间分布及组成分析。进行超微结构、组织化学和生化分析。对柱头乳突基部分泌物和柱头乳突表面及相邻假乳突之间中性脂质积累的检测表明,向日葵柱头表面是半干性的。以鲜重计,花粉被脂含量(8%)高于柱头(2.2%)。尼罗河红色荧光中性脂优先定位于花粉外壳。中性酯和三酰基甘油(TAGs)分别是花粉粒和柱头的主要脂质成分。木质素酸(24:0)和顺-11-二十烯酸(20:1)仅在花粉外壳中特异性表达。类似的长链脂肪酸在柱头表面花粉水化的初始信号机制中发挥了重要作用。脂肪酶(EC 3.1.1.3)活性在花粉粒和柱头中均有表达。柱头表达的酰基酯水解酶(EC 3.1.1.1)活性高于两个花粉组分。研究发现,花粉外壳特异性表达了两种酰基酯水解酶(41和38 kDa)。讨论了木质素酸(24:0)在花粉外壳中的特异性表达以及花粉和柱头中脂肪酶的定位,以确定它们在花粉-柱头相互作用中可能发挥的作用。
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引用次数: 23
The regulation of vesicle trafficking by small GTPases and phospholipids during pollen tube growth. 花粉管生长过程中小gtp酶和磷脂对囊泡运输的调控。
Pub Date : 2010-06-01 Epub Date: 2009-11-07 DOI: 10.1007/s00497-009-0118-z
Yan Zhang, Sheila McCormick

Polarized and directional growth of pollen tubes is the only means by which immotile sperm of flowering plants reach the deeply embedded female gametes for fertilization. Vesicle trafficking is among the most critical cellular activities for pollen tube growth. Vesicle trafficking maintains membrane homeostasis during rapid tube growth and provides polarity information by regulating protein/lipid compositions of different membrane compartments. In this review, we will focus on two classes of factors that orchestrate vesicle trafficking, small GTPases and phospholipids. We discuss the features of small GTPases and phospholipids that make them ideal components to regulate vesicle trafficking, review recent advances in understanding their involvement in vesicle trafficking, and propose directions for future research.

花粉管的极化和定向生长是开花植物的不动精子到达深埋的雌配子进行受精的唯一途径。囊泡运输是花粉管生长中最关键的细胞活动之一。在试管快速生长过程中,囊泡运输维持膜稳态,并通过调节不同膜室的蛋白质/脂质组成提供极性信息。在这篇综述中,我们将重点关注两类协调囊泡运输的因素,小gtp酶和磷脂。我们讨论了小gtp酶和磷脂的特点,使它们成为调节囊泡运输的理想成分,回顾了了解它们参与囊泡运输的最新进展,并提出了未来的研究方向。
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引用次数: 49
Semi-viviparous embryo development and dehydrin expression in the mangrove Rhizophora mucronata Lam. 红树半胎生胚胎发育及脱氢酶的表达。
Pub Date : 2010-06-01 Epub Date: 2010-01-19 DOI: 10.1007/s00497-009-0127-y
Flora Abdulrahman Ismail, Lisette M C Nitsch, Mieke M C Wolters-Arts, Celestina Mariani, Jan W M Derksen

Rhizophora mucronata Lam. is a tropical mangrove with semi-viviparous (cotyledon body protrusion before shedding), non-quiescent and non-desiccating (recalcitrant) seeds. As recalcitrance has been thought to relate to the absence of desiccation-related proteins such as dehydrins, we for the first time systematically described and classified embryogenesis in R. mucronata and assessed the presence of dehydrin-like proteins. Embryogenesis largely follows the classic pattern till stage eight, the torpedo stage, with the formation of a cotyledonary body. Ovule and embryo express radical adaptations to semi-vivipary in the saline environment: (1) A large, highly vacuolated and persistent endosperm without noticeable food reserves that envelopes the developing embryo. (2) Absence of vascular tissue connections between embryo and maternal tissue, but, instead, transfer layers in between endosperm and integument and endosperm and embryo. Dehydrin-like proteins (55-65 kDa) were detected by the Western analysis, in the ovules till stage 10 when the integuments are dehisced. An additional 50 kDa band was detected at stages 6-8. Together these results suggest a continuous flow of water with nutrients from the integument via the endosperm to the embryo, circumventing the vascular route and probably suppressing the initially induced dehydrin expression.

根霉;是一种热带红树林,具有半胎生(脱落前子叶体突出),不静止和不干燥(顽固)的种子。由于顽固性一直被认为与缺乏脱水相关的蛋白质(如脱氢蛋白)有关,我们首次系统地描述和分类了红毛鼠的胚胎发生,并评估了脱氢蛋白样蛋白质的存在。胚胎发生在很大程度上遵循经典模式,直到第八阶段,鱼雷阶段,形成子叶体。胚珠和胚胎在盐水环境中表达对半胎生的激进适应:(1)一个大的、高度空泡化的、持久的胚乳,没有明显的食物储备,包裹着发育中的胚胎。(2)胚与母体组织间无维管组织连接,胚乳与被皮、胚乳与胚间有转移层。Western分析法检测到脱氢蛋白样蛋白(Dehydrin-like protein, 55-65 kDa)存在于胚珠中,直至胚珠第10期被珠开裂。在6-8阶段检测到额外的50 kDa条带。综上所述,这些结果表明,水和营养物质通过胚乳从被皮源源不断地流向胚胎,绕过了维管通道,可能抑制了最初诱导的脱氢酶表达。
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引用次数: 13
Variability of the self-incompatibility reaction in Brassica oleracea L. with S 15 haplotype. 甘蓝s15单倍型自交不亲和反应的变异。
Pub Date : 2010-06-01 Epub Date: 2009-11-15 DOI: 10.1007/s00497-009-0119-y
Houria Hadj-Arab, Anne-Marie Chèvre, Thierry Gaude, Véronique Chable

Self-incompatibility (SI) is thought to have played a key role in the evolution of species as it promotes their outcrossing through the recognition and rejection of self-pollen grains. In most species, SI is under the control of a complex, multiallelic S-locus. The recognition system is associated with quantitative variations of the strength of the SI reaction; the origin of these variations is still not elucidated. To define the genetic regulations involved, we studied the variability of the SI response in homozygous S 15 S 15 plants in cauliflower. These plants were obtained from a self-progeny of a self-compatible (SC) plant heterozygous for S 15, which was generated after five selfing generations from one strongly self-incompatible initial plant. We found a continuous phenotypic variation for SI response in the offspring plants homozygous for the S 15 haplotype, from the strict SI reaction to self-compatibility, with a great proportion of the plants being partially self-compatible (PSC). Decrease in SI levels was also observed during the life of the flower. The number of pollen tubes passing through the stigma barrier was higher when counted 3 or 5 days after pollination than one day after pollination. Analysis of the expression of the two key genes regulating self-pollen recognition in cauliflower, the S-locus receptor kinase (SRK) and S-locus cysteine-rich (SCR/SP11) genes, revealed that self-compatibility or PSC was associated with decreased SRK or SCR/SP11 expression. Our work shows the particularly high level of phenotypic plasticity of the SI response associated with certain S-haplotypes in cauliflower.

自交不亲和(SI)被认为在物种进化中发挥了关键作用,因为它通过识别和排斥自交花粉粒来促进物种的异交。在大多数物种中,SI是由一个复杂的多等位基因s位点控制的。识别系统与SI反应强度的定量变化有关;这些变异的起源仍未阐明。为了明确相关的遗传调控,我们研究了花椰菜S - 15 S - 15纯合子植株的SI响应变异性。这些植株来自s15的自交亲和(SC)植株杂合的自交后代,该自交亲和植株由一株强自交亲和的初始植株自交5代后产生。我们发现,在s15单倍型纯合的后代植株中,从严格的SI反应到自亲和,SI反应存在连续的表型变化,大部分植株是部分自亲和(PSC)。在花的整个生命周期中,SI水平也呈下降趋势。通过柱头屏障的花粉管数在授粉后第3天或第5天比授粉后第1天要多。通过对花椰菜中调节自花粉识别的两个关键基因——s -座受体激酶(SRK)和s -座富含半胱氨酸(SCR/SP11)基因的表达分析,发现自亲和性或PSC与SRK或SCR/SP11的表达降低有关。我们的研究表明,花椰菜中与某些s单倍型相关的SI反应具有特别高的表型可塑性。
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引用次数: 13
Disruption of endosperm development: an inbreeding effect in almond (Prunus dulcis). 杏仁(Prunus dulcis)胚乳发育的破坏:近交效应。
Pub Date : 2010-06-01 Epub Date: 2009-10-21 DOI: 10.1007/s00497-009-0117-0
Encarnación Ortega, Pedro J Martínez-García, Federico Dicenta, José Egea

A homozygous self-compatible almond, originated from self-fertilization of a self-compatible genotype and producing a reasonable yield following open pollination, exhibited a very high fruit drop rate when self-pollinated. To investigate whether fruit dropping in this individual is related to an abnormal development of the embryo sac following self-fertilization, histological sections of ovaries from self and cross-pollinated flowers were observed by light microscopy. Additionally, the presence of pollen tubes in the ovary and fruit set were determined for both types of pollination. Despite pollen tubes reached the ovary after both pollinations, differences in embryo sac and endosperm development after fertilization were found. Thus, while for cross-fertilized ovules a pro-embryo and an endosperm with abundant nuclei were generally observed, most self-fertilized ovules remained in a previous developmental stage in which the embryo sac was not elongated and endosperm nuclei were absent. Although 30 days after pollination fruit set was similar for both pollination types, at 60 days it was significantly reduced for self-pollination. These results provide evidence that the high fruit drop in this genotype is the consequence of a disrupted development of the endosperm, what could be an expression of its high level of inbreeding.

一株纯合子自交亲和杏仁,起源于自交亲和基因型的自交亲和,在开放授粉后产量合理,自交亲和后的果实掉落率很高。为了研究该个体的果实掉落是否与自花受精后胚囊的异常发育有关,我们在光学显微镜下观察了自花和异花授粉花的卵巢组织学切片。此外,两种授粉方式的子房和坐果均存在花粉管。尽管两次授粉后花粉管都到达子房,但受精后胚囊和胚乳发育存在差异。因此,对于异交受精的胚珠,通常观察到原胚和具有丰富细胞核的胚乳,而大多数自交受精的胚珠仍处于胚囊未伸长和胚乳核缺失的先前发育阶段。尽管授粉后30天两种授粉类型的坐果量相似,但在60天自花授粉的坐果量显著减少。这些结果提供了证据,表明这种基因型的高落果率是胚乳发育中断的结果,这可能是其高水平近交的表现。
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引用次数: 10
Polycomb group gene function in sexual and asexual seed development in angiosperms. 多梳群基因在被子植物有性和无性种子发育中的作用。
Pub Date : 2010-06-01 Epub Date: 2009-12-29 DOI: 10.1007/s00497-009-0131-2
Julio C M Rodrigues, Ming Luo, Frédéric Berger, Anna M G Koltunow

In sexually reproducing angiosperms, double fertilization initiates seed development, giving rise to two fertilization products, the embryo and the endosperm. In the endosperm, a terminal nutritive tissue that supports embryo growth, certain genes are expressed differentially depending on their parental origin, and this genomic imbalance is required for proper seed formation. This parent-of-origin effect on gene expression, called genomic imprinting, is controlled epigenetically through histone modifications and DNA methylation. In the sexual model plant Arabidopsis, the Polycomb group (PcG) genes of the plant Fertilization Independent Seed (FIS)-class control genomic imprinting by specifically silencing maternal or paternal target alleles through histone modifications. Mutations in FIS genes can lead to a bypass in the requirement of fertilization for the initiation of endosperm development and seed abortion. In this review, we discuss the role of the FIS complex in establishing and maintaining genomic imprinting, focusing on recent advances in elucidating the expression and function of FIS-related genes in maize, rice, and Hieracium, and particularly including apomictic Hieracium species that do not require paternal contribution and thus form seeds asexually. Surprisingly, not all FIS-mediated functions described in Arabidopsis are conserved. However, the function of some PcG components are required for viable seed formation in seeds formed via sexual and asexual processes (apomixis) in Hieracium, suggesting a conservation of the seed viability function in some eudicots.

在有性繁殖的被子植物中,双受精启动种子发育,产生两个受精产物,胚胎和胚乳。胚乳是一种支持胚胎生长的末端营养组织,在胚乳中,某些基因的表达因亲本来源而异,这种基因组失衡是种子形成所必需的。这种亲本对基因表达的影响,被称为基因组印记,是通过组蛋白修饰和DNA甲基化来控制表观遗传的。在性模式植物拟南芥中,植物受精独立种子(FIS)类的Polycomb组(PcG)基因通过组蛋白修饰特异性沉默母本或父本靶等位基因来控制基因组印记。FIS基因的突变可导致胚乳发育起始和种子流产所需的受精绕过。在这篇综述中,我们讨论了FIS复合物在建立和维持基因组印记中的作用,重点介绍了最近在阐明玉米、水稻和Hieracium中FIS相关基因的表达和功能方面的进展,特别是包括不需要父系贡献从而无性形成种子的无融合Hieracium物种。令人惊讶的是,并非所有在拟南芥中描述的fis介导的功能都是保守的。然而,一些PcG组分的功能是在猴头属植物通过有性和无性过程(无融合)形成的种子中形成有活力的种子所必需的,这表明在一些异株植物中种子活力功能是保存的。
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引用次数: 46
期刊
Sexual Plant Reproduction
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