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1.
利用RNAi干扰技术研究不同基因对花粉发育、卵细胞发育和合子胚发育的影响是一种重要的手段。本研究通过筛选水稻在生殖发育过程中的9个重要调控基因,构建基因的RNAi表达载体,分析转基因植株育性及相关性状表型,以期探明RNAi表达载体对靶标性状的干扰效应。其中,AT61~AT63、AT64~AT66、AT67~AT69表达载体分别靶标花粉育性、卵细胞发育以及合子胚发育的调控。结果表明,除RNAi表达载体AT64没有获得转基因植株外,其余8个RNAi表达载体均获得了转基因植株;对T0代转基因植株的花粉育性、结实率以及潮霉素筛选(40 mg/L)发芽率检测的统计结果显示,RNAi表达载体AT62(花粉发育调控)、AT65(卵细胞发育调控)和AT67(合子胚发育调控)的干扰效应较强。本研究结果将为创制新型水稻基因工程不育系提供策略和选择。 相似文献
2.
棉花雄性不育研究和应用进展 总被引：7，自引：4，他引：7
综述了棉花雄性不育类型、不育机理和雄性不育系杂种优势利用状况,着重讨论了棉花雄性不育的细胞学、生理生化和分子生物学研究进展。并对本领域研究和应用中存在的问题及发展前景提出了看法。 相似文献
3.
植物花粉发育过程极其复杂,众多基因参与其中并对花粉发育的正常进行发挥着重要作用。植物花粉发育的基因工程研究是将花粉发育的分子水平研究和植物个体发育调控研究联系起来的中间环节,有助于全面认识植物花粉发育的机理。综述了植物花粉发育相关基因的分离、表达及其调控的研究进展,探讨了利用基因工程创造雄性不育的几种方法,并对今后的应用前景进行了展望。 相似文献
4.
为探讨反义寡聚核苷酸抑制(antisense oligodeoxynucleotides inhibition)技术在植物材料上的应用, 以已知功能的NtGNL1 (Nicotiana tabacum GNOM-Like1)为目标基因, 寻找反义寡聚核苷酸抑制最适作用体系, 并进一步分析NtGNL1的具体作用。根据目标基因mRNA序列设计反义寡聚核苷酸序列, 商业合成后并将其添加到烟草胚珠、种子和花粉管离体培养的培养基中, 以抑制NtGNL1的表达。结果表明, 将反义寡聚核苷酸引入离体培养系统, 短时间内抑制了目标基因mRNA的表达, 但对胚胎发育过程的影响和种子萌发的抑制都不明显。在花粉离体萌发系统中, 反义寡聚核苷酸抑制能够高效地引起目标基因mRNA的下调。显微缩时对FM4-64染色的花粉管观察发现, NtGNL1的下调能够引起囊泡分布和运输方向的改变。对花粉管膜流相关的5个基因的半定量分析也显示反义寡聚核苷酸进入花粉管后, 导致3个基因的表达下调, 暗示NtGNL1抑制表达会影响花粉管囊泡运输的多个节点。 相似文献
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6.
高粱雄性不育系、保持系正负极向过氧化物酶同工酶的电泳比较 总被引：2，自引：0，他引：2
对高粱雄性不育系、保持系不同生长发育阶段叶片中的正、负极向过氧化物酶同工酶进行了检测。结果表明,叶片中正极向过氧化物酶同工酶谱两者基本相同,说明正极向过氧化物酶在不同生长发育时期是稳定的。负极区域中,在叶片发育过程中有2条酶带消失,证明在这个区域中遗传信息出现顺序表达现象。花粉粒发育过程中,不育系的正极向过氧化物酶带始终多于保持系。作者认为,不育细胞质基因可影响编码过氧化物酶的核基因的表达,引起雄性不育。不育细胞质基因可能也同时具有调节基因的功能,表现为具有解除阻遏的作用,使不育系花粉粒发育过程中部分过氧化物酶基因得以表达;而保持系不具有这种解除阻遏的细胞质不育基因,使部分基因仍处于阻遏状态,造成保持系中过氧化物酶同工酶谱带少于不育系的现象。不育细胞质基因的这种调控作用,同样有可能影响核基因遗传功能的正常发挥,引起核质之间的不协调而导致雄性不育。 相似文献
7.
The tomato floral system is distinguished from that of Arabidopsis and Antirrhinum in several ways. The shoot is a sympodium, the first flower of the inflorescence is apical and no bracts are formed. We discuss the possible function of genes affecting growth habit and inflorescence development and analyze in detail the unique developmental alterations caused by inhibition or ectopic expression of tomato-specific MADS-box genes. Preliminary analysis of transgenic tomato plants expressing an alien homeotic gene and several different chimeric genes of the MADS-box family is also reported. 相似文献
8.
PEI Wen-Feng WU Jian-Yong XING Chao-Zhu GUO Li-Ping QI Ting-Xiang WANG Hai-Lin 《棉花学报》2013,25(6):478-485
This study aimed to analyze the expression level and relationship between miR156 and its target gene TBCC (Tubulin binding Cofactor C) in buds of Gossypium at different developmental stage in a cytoplasmic male sterile(CMS) line and its maintainer line, to clone the full-length cDNA of the TBCC gene, and to determine the possible role of the TBCC gene in pollen development. Real-time RT-PCR showed that the expression levels of miR156 gradually increased with the development of pollen between the CMS line and the maintainer line. The expression of GhTBCC was strikingly complementary with the expression of miR156 in the maintainer line, but showed very low expression levels in the CMS line, which demonstrated that TBCC is related to male sterility. The full-length cDNA sequence of TBCC was cloned from the buds of the maintainer line, and the gene was named as GhTBCC. The CDS of GhTBCC is 1713 bp and encodes a predicted protein of 570 amino acids with a predicted molecular weight of 62.79 kDa. Conserved TBCC and CARP domains were observed in the predicted protein sequence. GhTBCC showed 81 %, 80 %, and 77 % sequence similarity to the TBCC protein sequences of Populous trichocarpa, Ricinus communis, and Arabidopsis thaliana, respectively. 相似文献
9.
棉花育性相关基因GhPG2的克隆与表达分析 总被引：2，自引：1，他引：1
多聚半乳糖醛酸酶家族是一个细胞壁水解酶,在许多高等植物的花粉中都具有高水平外切多聚半乳糖醛酸酶(PG)活性,可能在花粉成熟和花粉管伸长中发挥重要作用。利用已报道与恢复基因连锁的分子标记STS679筛选陆地棉Y18恢复系核基因组BAC文库,获得一个阳性克隆Z55E7,对该克隆测序获得6个基因,其中一个为编码多聚半乳糖醛酸酶(polygalacturonase)基因,暂被命名为GhPG2。在棉花细胞质雄性不育系、保持系和恢复系中对该基因进行序列和表达分析,以便进一步了解该基因与棉花细胞质雄性不育的关系。从BAC克隆中分离到GhPG2基因,对其进行序列分析,以半定量RT-PCR和实时定量PCR分析该GhPG2基因在棉花恢复系、保持系和不育系各个组织中的表达模式。分离到GhPG2基因的基因组序列为1 518 bp,其开放阅读框序列为1 239 bp,共编码412个氨基酸,包含PG基因家族的4个保守结构域。系统进化树分析表明,GhPG2基因与其他物种花粉表达的PG基因同属C分支。表达分析表明, 该基因在恢复系和保持系的花药、花瓣和VI级花蕾中高表达,而不育系的相应组织该基因表达量明显降低。GhPG2基因归类于花粉表达的一类PG基因。实验结果暗示GhPG2基因可能与棉花正常的花器官发育相关。 相似文献
10.
棉花耐盐相关基因GhVP的表达及功能分析 总被引：3，自引：2，他引：1
为了研究Gh VP基因在棉花中的表达特性和功能,克隆了棉花Gh VP基因,构建p BI121-Gh VP∷GFP荧光表达载体,采用基因枪技术转化洋葱内表皮细胞,结果显示Gh VP基因编码蛋白定位于细胞质膜上。将构建的p BI121-Gh VP∷GFP荧光表达载体转化棉花花粉,花粉荧光明显增强,说明该基因可以在棉花中表达,为Gh VP基因可以在棉花中表达提供了依据,为培育棉花耐盐新材料奠定了基础。 相似文献
11.
K. K. Nkongolo 《Euphytica》1996,90(3):337-344
Summary The Barley Yellow Dwarf Virus disease (BYDV) and the Russian wheat aphid (RWA) Diuraphis noxia (Mordvilko) have caused significant losses to wheat and barley in several areas of the world. Important sources of resistance to both BYDV and RWA have been found in Triticale. Different generations of interspecific wheat x Triticale crosses were produced and the progenies were screened for BYDV and RWA tolerance. Plants with equal chromosome numbers showed different levels of fertility. A significant correlation was observed between pollen fertility and seed set in primary florets (r=0.57). In generaL, pollen fertility, seed set and the number of euploid plants (2n=42) increased from one generation to the next. The expression of BYDV tolerance varied from population to population. Additive effects were predominant in F1 and some backcross populations. A dominant effect of rye tolerance genes was also observed in few populations. A monogenic trait or a quantitative (polygenic) character would not agree with the observed segregation patterns. The heritability of this oligogenic tolerance was quite different between populations and in many populations the tolerance genes were only partially expressed. Some transgressive segregation for tolerance and sensitivity was demonstrated. The genes controlling tolerance to RWA in Triticale lines, Muskox 658 and Nord Kivu were not expressed in advanced lines resistant to BYDV. This indicates that tolerance genes for BYDV and RWA in these lines are located on different chromosomes. 相似文献
12.
Summary Isozyme expression of malate dehydrogenase (MDH), phosphoglucomutase (PGM), and phosphoglucose isomerase (PGI) were examined from 12 tissues of pecan by starch gel electrophoresis. Tissue type, stage of development, and extraction method had little effect on isozyme expression of MDH or PGM but did influence PGI expression. Additional PGI bands were observed from seed tissue and soaked pollen. A direct relationship was observed between PGI expression from seed tissue and eventual expression from the germinated seedling. Additional PGI bands from soaked pollen were thought to be produced from structural changes and not due to more efficient extraction. The genotype of immature pecan seeds could be determined for Mdh-1, Pgi-2, and Pgm-1 provided cellular endosperm had developed. Watery endosperm did not contain detectable levels of the study enzymes. The advantages of uniform isozyme expression among various tissues and ability to determine the genotype of immature pecan seed are discussed. 相似文献
13.
Summary Selection among microgametophytes usually exploits variation in pollen grain germination. Studies of variation in pollen grain size in common bean (Phaseolus vulgaris L.) suggested that selection for size might lead to changes in sporophytic traits. To determine whether microgametophytic selection based on size would affect pollen grain size in subsequent generations or sporophytic traits that were correlated with pollen grain size, pollen grains from three crosses were separated into two size categories by sieving and then used to pollinate cv. Diacol Calima. Selection resulted in changes in pollen grain diameter for pollen from F1, F2 and F3 plants for all crosses. In vitro germination indicated no differences between vigor of large and small grains, but extraction and sieving reduced germinability. F1 seed from two of the crosses with size-selected pollen varied in weight according to pollen grain size, but in subsequent generations, the effect disappeared. Both size categories of selected pollen resulted in F2 progeny with reduced numbers of seeds per pod as compared to controls, suggesting that the size selection process may have resulted in indirect selection for traits reducing seed set. The overall results suggested that genes determining pollen grain size in bean have little or no effect on sporophytic traits such as seed size and seed yield. 相似文献
14.
为了寻找小麦单核中晚期愈伤组织诱导率较高的原因,探讨小麦花药发育的蛋白质代谢机制,本试验利用双向电泳技术对陕农138小麦小孢子单核中晚期和双核期的全蛋白分析表明,在等电点4~7之间可识别约450个以上清晰的蛋白质点,检测到差异点26个,对其中14个质量较好、重复性较高的差异表达的蛋白质点采用基质辅助激光解吸分离飞行时间质谱(MALDI-TOF-MS)进行肽指纹图谱分析,并利用Mascot软件在NCBInr数据库搜索,鉴定出11个蛋白质点,另3个蛋白质点未得到有意义的鉴定,11个蛋白质点分别被鉴定为UDP-葡萄糖焦磷酸化酶(1个蛋白点)、叶绿素抗体结合蛋白(1个蛋白点)、pentatricopeptide重复蛋白PPR566-6 (1个蛋白点)、半胱氨酸蛋白酶抑制剂(1个蛋白点)、泛素(1个蛋白点)、S-腺苷-L-半胱氨酸水解酶(2个蛋白点)、放氧增强蛋白(2个蛋白点)和假定蛋白(2个蛋白点)。这些蛋白质点的功能涉及到糖代谢、蛋白质降解、细胞防卫等代谢过程。 相似文献
15.
本研究目的在于利用化学杀雄剂"化杀灵WP1"诱导甘蓝型油菜R121产生雄性不育,并对花蕾中的差异表达基因进行分离、筛选及定量表达分析。首先,通过抑制消减杂交技术(SSH)和反向Northern斑点杂交技术,分离和筛选出在雄性不育花蕾中差异表达的基因片段。然后,将差异片段测序后在NCBI数据库中进行同源性比对及功能分析。最后,对差异表达基因进行反转录荧光定量表达(RT-QPCR)分析。本研究从筛选出来的6个阳性克隆中获得一个与多聚半乳糖醛酸酶基因(MF6)100%同源的基因片段,该基因被认为与花粉生长发育及花粉成熟密切相关。对MF6基因在不同长度及不同育性花蕾中的表达量进行了定量分析,结果显示,MF6基因在雄性可育花蕾中的表达量均高于雄性不育花蕾,且在蕾长1.00mm～1.50mm和3.50mm～4.00mm时分别达到了18.53倍和43.15倍的差异。因此,我们推测"化杀灵WP1"主要通过抑制花粉母细胞减数分裂时期中MF6基因的表达,使花粉产生败育;同时,MF6基因的表达可能也与花丝的伸长生长有关。 相似文献
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一个棉花β-1,3-葡聚糖酶基因全长cDNA的克隆与特征分析 总被引：1，自引：1，他引：0
利用本实验室分离的防卫基因类似物片段,通过5’和3’RACE,从海岛棉海7124中克隆了一个β-1,3-葡聚糖酶基因全长cDNA,命名为GbGLU。其全长1 266 bp,含一个编码361个氨基酸残基的开放阅读框。该基因编码产物理论分子量和等电点为MW = 39.66 kD,pI = 9.15,含有糖基水解酶家族17的保守结构域,N端有26个氨基酸残基组成的信号肽。Southern结果显示,GbGLU在棉花基因组中以单拷贝形式存在,研究发现,GbGLU的表达受黄萎病菌的诱导。聚类分析表明,棉花葡聚糖酶基因可分为3类,分别含有糖基水解酶家族3、9、17的保守结构域。无论棉花中的还是本研究中参考的其他作物中,受病菌诱导的葡聚糖酶基因都含有糖基水解酶家族17的保守结构域,故推断其为此类基因参与抗病反应的功能域。 相似文献
17.
小麦雄性不育系保持系过氧化物酶同工酶的比较研究 总被引：2，自引：0，他引：2
从小麦雄性不育系、保持系的10个不同组织、器官中共检测出8种过氧化物酶,在叶、芽鞘、根、节、节间、穗轴、叶鞘和芒中,不育系和保持系的过氧化物酶基本相同,说明这种酶由细胞核编码,但在花粉粒发育的3个不同时期,不育系与保持系间的过氧化物酶有明显差异,表现为不育系中的过氧化物酶同工酶多于保持系,这可能是由于不育系中细胞质不育基因对核基因的表达有调控作用,这种调控作用表现为不育细胞质基因的去阻遏作用,即不育细胞质基因可以去除细胞核中编码过氧化物酶基因的阻遏作用,使不育系中有更多的过氧化物酶基因得以表达,造成不育系中过氧化物酶同工酶多于保持系的现象. 相似文献
18.
Most wild plants and some crops of the Brassicaceae express self-incompatibility, which is a mechanism that allows stigmas to recognize and discriminate against “self” pollen, thus preventing self-fertilization and inbreeding. Self-incompatibility in this family is controlled by a single S locus containing two multiallelic genes that encode the stigma-expressed S-locus receptor kinase and its pollen coat-localized ligand, the S-locus cysteine-rich protein. Physical interaction between receptor and ligand encoded in the same S locus activates the receptor and triggers a signaling cascade that results in inhibition of “self” pollen. Sequence information for many S-locus haplotypes in Brassica species has spurred studies of dominance relationships between S haplotypes and of S-locus structure, as well as the development of methods for S genotyping. Furthermore, molecular genetic studies have begun to identify genes that encode putative components of the self-incompatibility signaling pathway. In parallel, standard genetic analysis and QTL analysis of the poorly understood interspecific incompatibility phenomenon have been initiated to identify genes responsible for the inhibition of pollen from other species by the stigma. Herewith, we review recent studies of self-incompatibility and interspecific incompatibility, and we propose a model in which a universal pollen-inhibition pathway is shared by these two incompatibility systems. 相似文献
19.
华北驼绒藜花粉萌发特性及不同授粉方式对其结实率的影响 总被引：1，自引：0，他引：1
为寻找华北驼绒藜结实率低、群落退化的原因,在栽培条件下对其花粉生活力、寿命、柱头萌发率与结实率的关系进行了相关性研究。采用同株、异株、自然授粉处理华北驼绒藜,同株授粉结实率(81.53%)显著低于异株授粉(88.6%)和自然授粉(92.6%)的结实率;采用TTC法对华北驼绒藜花粉寿命和生活力的研究结果表明,其平均寿命为6d,最佳授粉时间为1~3d,生活力达95%左右,不存在花粉败育和短命的问题,即不存在小孢子发育障碍。3种授粉方式处理后,采用荧光显微镜观察华北驼绒藜花粉在柱头上的萌发率,萌发高峰集中在授粉后3~4h,萌发率达90%以上,三者不存在显著差异,即华北驼绒藜不存在花粉与柱头的识别障碍。结实率与花粉在柱头萌发率的相关分析结果表明,二者不存在相关性。研究初步认为,同株授粉产生结实率低的原因可能发生在授精或胚胎发育过程,有待进一步研究。目前,华北驼绒藜野生群落严重退化,植被密度减少,可能存在大量自株授粉是造成群落退化的原因之一。 相似文献
20.
Nicky Driedonks Mieke Wolters-Arts Heidrun Huber Gert-Jan de Boer Wim Vriezen Celestina Mariani Ivo Rieu 《Euphytica》2018,214(4):67
Climate change has become a serious threat for crop productivity worldwide. The increased frequency of heat waves strongly affects reproductive success and thus yield for many crop species, implying that breeding for thermotolerant cultivars is critical for food security. Insight into the genetic architecture of reproductive heat tolerance contributes to our fundamental understanding of the stress sensitivity of this process and at the same time may have applied value. In the case of tomato (Solanum lycopersicum), germplasm screenings for thermotolerance have often used yield as the main measured trait. However, due to the complex nature of yield and the relatively narrow genetic variation present in the cultivated germplasm screened, there has been limited progress in understanding the genetic basis of reproductive heat tolerance. Extending the screening to wild accessions of related species that cover a range of climatic conditions might be an effective approach to find novel, more tolerant genetic resources. The purpose of this study was to provide insight into the sensitivity of individual reproductive key traits (i.e. the number of pollen per flower, pollen viability and style protrusion) to heat-wave like long-term mild heat (LTMH), and determine the extent to which genetic variation exists for these traits among wild tomato species. We found that these traits were highly variable among the screened accessions. Although no overall thermotolerant species were identified, several S. pimpinellifolium individuals outperformed the best performing cultivar in terms of pollen viability under LTMH. Furthermore, we reveal that there has been local adaptation of reproductive heat tolerance, as accessions from lower elevations and higher annual temperature are more likely to show high pollen viability under LTMH. 相似文献