苹果花药培养不定胚形成的细胞学观察  

A cytological observation on the development of embryoid in cultured anther of apple

作　　者：聂园军 冯丽云[2] 张春芬[3] 邓舒[3] 肖蓉[3] 侯丽媛[4] 赵菁 曹秋芬[4] NIE Yuanjun;FENG Liyun;ZHANG Chunfen;DENG Shu;XIAO Rong;HOU Liyuan;ZHAO Jing;CAO Qiufen(Agricultural Resource and Economic Research Institute,Shanxi Academy of Agriculture Sciences,Taiyuan 030031,Shanxi,China;Millet Research Institute Shanxi Academy of Agricultural Sciences,Changzhi 046011,Shanxi,China;Pomology Institute,Shanxi Academy of Agricultural Sciences Taiyuan 030031,Shanxi,China;Biotechnology Research Center,Shanxi Academy of Agriculture Sciences,Taiyuan 030031,Shanxi,China)

机构地区：[1]山西省农业科学院农业资源与经济研究所,太原030031 [2]山西省农业科学院谷子研究所,山西长治046011 [3]山西省农业科学院果树研究所,太原030031 [4]山西省农业科学院生物技术研究中心,太原030031

出　　处：《果树学报》2020年第3期322-329,共8页Journal of Fruit Science

基　　金：山西省重点研发-国际合作项目(201803D421023);山西省青年科技研究基金(201701D221168);国家自然科学基金项目(31372033);山西省农业科学院优势课题组项目(YCX2018DZYS15);山西省科技成果转化与引导专项(201904D131039)。

摘　　要：【目的】通过对苹果花药离体培养过程中小孢子的发生及雄核发育的过程进行了系统的细胞学观察,明确花粉培养不定胚形成的关键时期和类型,为苹果花药培养小孢子不定胚形成的机理研究奠定基础。【方法】以‘嘎拉’苹果不同发育时期的花蕾为试材,采用石蜡切片技术对其小孢子发生和雄配子发育过程进行了解剖学观察,并对接种培养后的花药进行了切片制作,系统观察了小孢子不定胚的发育过程。【结果】单核期花粉经均等分裂后形成两个对称的两核细胞,然后持续多次均等分裂后形成多细胞(核)结构,进一步分化发育成不定胚。【结论】为苹果花药所获植物的单倍体来源提供了直接证明,有关苹果雄核发育的机理研究还需要通过小孢子培养进一步研究。【Objective】Anther culture is an important method to produce haploid from microspores,and is widely used in plant breeding research.Apple is a temperate fruit species which is characterized by a long reproductive cycle with a juvenile phase of several years,a large tree size and a high degree of heterozygosity derived from self-incompatibility.It is almost impossible to obtain a homozygous plant by the conventional hybrid breeding.Production of haploids by anther culture offers new possibilities for genetic studies and breeding.Because of presence of anther wall cells,the regenerated plants via anther culture often have different levels of polidy,such as haploids,diploids,triploids,tetraploids,and chimeras.Therefore,homozygous analysis must be conducted to confirm whether regenerated buds are derived from pollen grains.Cytological oberservations on the development of embryoids inside the cultured anthers is a direct evidence to prove that the plants are developed from the microspores.At the same time,embryoid induction procedures in anther culture of apple will be clarified at the cellular level.【Methods】The cultivar of‘Gala’was used in this paper.In early April,the full flower buds were selected from different directions in the upper and outer parts of the canopy and the buds of different developmental stages were used to observe the occurrence of microspores and the development of androgenesis.The cultured anthers were used to study the process of embryoids development.Samples were taken at 0,5,10,12,15,17,20,25,30,40,50 and 60 days after inoculation.Then the flower buds and anthers at each period were fixed by FAA for 12-24 hours,and the conventional paraffin sections were obtained after series alcohol dehydration.The 6-8 m thick sections were double stained with hematoxylin and saffron.Morphological characteristics of anthers and their internal microsporocytes in different periods were observed under the light microscope.【Results】Based on the cytological oberservations,development of microspore

关 键 词：苹果 花药 不定胚 小孢子发生 雄核发育 

分 类 号：S661.1[农业科学—果树学]