作者：唐毓玮 李佳慧 毛立彦 黄秋伟 龙凌云 於艳萍 苏
来源：《热带作物学报》2022年第04期
摘 要：睡蓮属（Nymphaea）植物是我国新兴的水生花卉，其中重瓣型睡莲的花态丰满艳丽，一直倍受睡莲爱好者、育种者的喜爱和关注，在水生园林景观中的应用日渐广泛，但在育种的过程中，育种者经常遇到杂交不结实的现象，导致重瓣型睡莲品种难以培育。为探索重瓣睡莲难以结实的原因，以重瓣睡莲中较具代表性的品种‘泰国王’为实验组，以育性较好的品种‘公牛眼’为对照组，利用石蜡切片技术对其授粉后不同时期的子房进行显微结构比较，采用扫描电镜进一步观察样本的柱头和胚珠，通过RNA-seq测序分析发育与败育子房的基因表达差异。结果表明：2个睡莲品种的柱头表面均分布着多细胞单列乳突，细胞间的连接处形成一圈圈凹槽，乳突数量庞大、排列紧密，配合自身的结构能更容易捕获外来的花粉。‘公牛眼’授粉7d后子房中大部分胚珠发育成红种子，10d后子房膨大成果实，种皮由红转黑，形成成熟的种子;重瓣睡莲‘泰国王’授粉7d后无种子形成，子房中有部分胚珠发育，其表皮变为红，形态特征与‘公牛眼’授粉4 d后的红胚珠相似，由此推测少量胚珠可能完成了受精，但无法进一步发育，10 d后子房完全败育。显微结构观察可知，授粉7d后2个品种的胚珠发育产生明显差异，‘公牛眼’的胚珠明显增大，合子开始分裂形成胚，而‘泰国王’胚珠内珠心萎缩，胚珠逐步败育消解，授粉10 d后，‘公牛眼’胚珠的珠被增厚形成种皮，胚开始发育，进一步向种子形态转变。由转录组测序分析可知，差异 基因显著富集在光合作用——天线蛋白、光合作用、卟啉和叶绿素代谢、核糖体、淀粉和蔗糖代谢等通路，这些通路可能是影响睡莲子房发育的关键途径，推测‘泰国王’子房败育可能是因为自身光合作用受到抑制，进而影响了蔗糖、淀粉等营养物质的生成。
帕拉米韦
关键词：睡莲;杂交授粉;子房发育;胚珠;转录组中图分类号：S682.32 文献标识码：A
Differences in Ovary Development Between Nymphaea ‘Bull’s Eye’ and Nymphaea ‘King of Siam’ after Pollination
TANG Yuwei LI Jiahui MAO Liyan HUANG Qiuwei LONG Lingyun YU Yanping SU Qun
1. Guangxi Subtropical Crop Research Institute， Nanning， Guangxi 530001， China; 2. Flower Research Institute， Guangxi Academy of Agricultural Sciences， Nanning， Guangxi 530007， China
Abstract： Nymphaea 热解焚烧炉（water lily） is an aquatic flower that has been introduced in recent years in our country. In particular， double petal water lily are very popular and f
ull of love and attention by water lily lovers and breeders. And it has been increasingly widely used in the field of aquatic garden landscapes. But in the process of breeding， breeders often encounter the situation of failed hybridization. This makes it hard for breeders to breed double flower. In order to study on the reason that double petal water lily is difficult to set fruit， we selected the more representative cultivar Nymphaea ‘King of Siam’ from double petal water lily as the experimental group， and the better fertile cultivar Nymphaea ‘Bull’s Eye’ as the control group. Firstly， the microstructure of ovaries at different stages after pollination was compared by paraffin section technique. Secondly， the stigmas and ovules were further observed by scanning electron microscope. Finally， the differences of gene expression between developing and abortive ovary were analyzed by RNA-Seq. The results showed that multicellular uniserial papillae were distributed on the stigma surface of the two water lily varieties， and a circle of grooves were formed at the junction between single cells. The number of papillae was large and closely arranged. We think this structure can more easily capture foreign pollen. Meanwhile， after 7 days of N. ‘Bull’s Eye’ pollination， most of the ovules
developed into red seeds， 10 days later， the ovary expanded into fruit， and the seed coat changed from red to black to form mature seeds. On the other hand， after 7 days of pollination， no seeds were formed in N天弩无人机. ‘King of Siam’. However， some ovules in its ovary develop， and its epidermis turns red. The morphological characteristics are similar to the red ovules after 4 days of N. ‘Bull’s Eye’ pollination. Therefore， we speculate that a small number of ovules may have completed fertilization， but can not develop further. Than， the ovary was completely aborted after 10 days. Microstructural observation showed that there were significant differences in ovule development between the two varieties after 7 days of pollination. The ovules of N江苏省物业管理条例. ‘Bull’s Eye’ increased significantly， and the zygotes began to form embryos， while the nucellus in the ovules of N. ‘King of Siam’ shrank and the ovules gradually disappeared. After 10 days of pollination， the embryo of N. ‘Bull’s Eye’ began to develop and further changed to seed morphology. In additional， the enrichment analysis of differential genes showed that photosynthesis antenna protein， photosynthesis， porphyrin and chlorophyll metabolism， ribosome， starch and sucrose metabolism may be the key pathways affe
cting the development of waterlily ovary. Consequently， we speculate that the abortion of the ovary of N. ‘King of Siam’ may be due to the inhibition of its own photosynthesis， which affects the production of nutrients such as sucrose and starch.
Keywords： Nymphaea; cross pollination; ovary development; ovule; transcriptome
DOI： 10.3969/j.issn.1000-2561.2022.04.019
授粉是種子植物生殖过程中的重要阶段[1]，授粉成功的子房会进一步发育成果实，而个体间不亲和的授粉会影响花粉与雌蕊的识别、受精以及胚珠的发育中国油脂网[2-4]，进而导致子房败育无法结实。
芥川龙之介河童 睡莲是睡莲科（Nymphaeaceae）睡莲属（Nymphaea）植物的通称[5]，花丰富，形态多样，在水生园林景观中应用广泛。国内育种研究始于1999年，育种目标以花特异性为主。截止2019年，我国已培育成新品种205个，其中国内鉴定18个、国际登陆187个[6]。重瓣型睡莲的花态丰满艳丽，一直倍受睡莲爱好者、育种者的喜爱和关注，但在育种的过程中，育种者经常遇到杂交不结实的现象[7-9]，限制了重瓣型热带睡莲的杂交育种研
究。
‘泰国王’是重瓣型热带睡莲中较具有代表性的品种，本课题组前期进行了杂交试验，发现‘泰国王’授粉以后常出现假膨大的现象，子房略微膨大后衰败，难以结实。本研究选择睡莲‘泰国王’品种和育性较好的‘公牛眼’品种为材料，采用组织学、转录组学等手段研究其授粉后子房发育的差异，探讨重瓣型睡莲授粉后子房败育的成因，以期为睡莲的杂交育种提供理论基础。
1.1 材料
实验材料为栽培多年的热带睡莲品种‘公牛眼’和‘泰国王’，种植于广西壮族自治区南宁市兴宁区亚热带作物研究所睡莲种质资源圃（108°38¢E，22°87¢N），实验地属于亚热带季风气候。课题组前期研究发现，‘泰国王’的雄蕊完全瓣化，无法产生花粉，因此不能进行自交授粉，仅能作为母本材料进行杂交，经人工杂交授粉后难以结实，而‘公牛眼’自交结实率较高。
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