密级:论文编号:
学位论文
设施蓝莓蜂授粉及不同授粉强度下
柱头转录组学研究
硕士研究生：冀薇
学号：82101212375
指导教师：姚军副研究员
申请学位类别：农学硕士
专业：特种经济动物饲养
研究方向：特种经济动物病虫害及其防治
培养单位：蜜蜂研究所
研究生院
2024年6月
Secrecy:No.
Chinese Academy of Agricultural Sciences
Thesis
Pollination of Blueberry in Greenhouse and
Transcriptomic Analysis of Stigma in Response to the
Intensity of Pollination
Candidate:JI Wei
Student ID:82101212375
Supervisor:Associate Prof.YAO Jun
Degree Type:Master of Science in The Rearing
of Special-type Economic Animals
Major:The Rearing of Special-type Economic
Animals
Research Field:Disease and Treatment of
Special Economic Animals
Institution:Institute of Apicultural Research
Graduate School
June 2024
摘要
蓝莓在全球范围内均有种植，随着设施栽培面积的不断扩大，为确保蓝莓的产量和品质，实
施充分、高效的授粉非常重要。授粉受精过程是蓝莓进行有性生殖的最初环节，这一过程的顺利
完成有赖于蓝莓自身的品种特点、传粉蜂的授粉以及蓝莓柱头与花粉间的相互作用。本文以设施
种植的‘蓝源（M7）’蓝莓为研究对象，首先通过对其开花生物学的研究，评估合适的授粉效期。
同时对设施蓝莓中常用的西方蜜蜂（Apis mellifera）、东方蜜蜂（Apis cerana）和地熊蜂（Bombus
terrestris）的访花行为和访花后柱头花粉沉降数进行观察和计数，评估不同传粉蜂的授粉效率。
由于不同授粉强度影响蓝莓的生殖与果实发育，为进一步探究蓝莓的响应机制及其表达特性，本
研究对3种授粉强度处理下的蓝莓柱头进行转录组测序和分析，并对不同授粉强度下蓝莓果实品
质的差异进行比较。主要研究结果如下：
（1）通过测定设施栽培的‘蓝源（M7）’开花生物学特性，发现其在开花第2天具备较优
的授粉特性。‘蓝源（M7）’的花粉为四合体复合花粉，平均每朵花的花粉总量为14,560±766.29
粒；随花龄的不断延长，‘蓝源（M7）’柱头可授性表现为先升高后逐渐降低的趋势，开花1~2d
可授性较强，持续至4d 左右；‘蓝源（M7）’在开花前没有花蜜产生，蓝莓花单日泌蜜量和单花
期累积泌蜜量在开花前两天内均呈上升趋势，在开花第2天花蜜的含糖量最高为42%。
（2）通过比较三种传粉蜂访花行为和授粉效率，发现就单只蜂而言，地熊蜂是比东方蜜蜂
和西方蜜蜂更有效的传粉者。三种传粉蜂的单次访花时间均存在显著差异（p <0.001），西方蜜蜂
的单次访花时间（18.55±0.44s）最长，东方蜜蜂（12.57±0.16s）次之，地熊蜂（9.31±0.41s）
最短；无蜂访问的蓝莓花柱头沉降数为14.65±2.520粒，单次访问后地熊蜂沉积的花粉量（190.90
±15.990）最高，其次是东方蜜蜂（152.98±15.880）和西方蜜蜂（142.70±15.170）；通过比较
全天开放传粉后的柱头花粉沉积量发现，地熊蜂沉积的花粉数量最多（286.09±23.225），三种蜂
的比较趋势与单次访问数据趋势一致。
（3）提高授粉强度对果实发育具有积极影响。设置5个授粉处理组，不同授粉强度下蓝莓
的果型指数和单粒重具有显著差异，随着授粉强度的增加，蓝莓坐果率、单果重、果实横径及果
实纵径呈现上升趋势。未授粉组及具活花粉占比0%组（F0组）均未发育为成熟果实；相较于另
两组，熊蜂一天内开放授粉组（BO 组）授粉的果实个体大，单粒较重。
（4）授粉强度的增加可能对柱头和花粉的识别过程产生影响，通过参与柱头和花粉间的信
号转导、激素合成过程对花粉管的生长起到促进作用。随着授粉强度的增加，参与花粉管生长调
节的钙依赖蛋白激酶17（Cpk17），涉及肌醇含量的肌醇加氧酶（MIOX1）等基因的表达量显著
提高。结合GO 功能注释和KEGG 通路富集，发现授粉强度提高可影响与细胞壁组织形成和肌醇
膦酸代谢相关的生物学功能变化，从而影响花粉管的生长。
关键词：蓝莓，授粉，效率，转录组，果实品质
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Abstract
Blueberries are extensively cultivated worldwide,with a growing focus on efficient and precise
pollination to enhance yield and quality.Pollination and fertilization are crucial for the sexual
reproduction of blueberries,relying on ecological traits,pollination by bees,and stigma-pollen
interactions.This study focused on the blueberry variety 'Bluesouth (M7)',evaluating the optimal
pollination period by studying its ecological characteristics.Foraging behaviors of Apis mellifera,Apis
cerana,and Bombus terrestris were monitored,along with stigma pollen deposition,to evaluate
pollinator efficiency.Different levels of pollination intensity were found to impact blueberry
reproduction and development.Transcriptome analysis of blueberry stigmas under varying pollination
intensities revealed insights into the plant's response mechanisms and gene expression patterns.
Additionally,comparisons of fruit quality under different pollination methods were conducted.The
main findings are as follows:
(1)Research on the flowering biology of blueberry indicates that as the flower age increases,the
stigma receptivity of 'Bluesouth (M7)'initially rises,then gradually declines.Stigma receptivity is
strongest after 1to 2days of flowering,lasting until approximately 4days.The average total amount of
pollen in each flower of 'Bluesouth (M7)'is 14,560±766.29pollen tetrads.Additionally,'Bluesouth
(M7)'flowers do not produce nectar before flowering,but as they age,the amount of nectar secreted
within 24hours and the total nectar secretion during a single flowering period increase.Moreover,the
sugar concentration of the nectar also increases with flower age,reaching its peak on the second day of
flowering.
(2)The study focused on the flower visiting behavior and pollination efficiency of pollinating bees,
it was found that B.terrestris is a more efficient pollinator compared to A.cerana and A.mellifera on an
individual level.Results indicated significant differences in the duration of flower visits among the three
pollinator bees (p <0.001),with A.mellifera having the longest visit duration (18.55±0.44s),followed
by A.cerana (12.57±0.16s)and B.terrestris (9.31±0.41s).In terms of stigma pollen deposition
number,blueberry flowers without bee visits had an average of 14.65±2.520pollen depositions.After a
single visit,B.terrestris deposited the highest amount of pollen (190.90±15.990),followed by A.
cerana (152.98±15.880)and A.mellifera (142.70±15.170).Consistent with single visit data,open
pollination throughout the day also showed B.terrestris depositing the highest amount of pollen (286.09
±23.225).
(3)Enhancing pollination intensity positively influences fruit development.In response to increased
pollination intensity,blueberry fruit setting rate,single fruit weight,fruit transverse diameter,and fruit
longitudinal diameter all exhibited an upward trend.Within one day,the fruits of B.terrestris open
pollination group were notably large in size.Conversely,neither the non-pollination group nor the F0
group successfully developed into mature fruits.
(4)Increased pollination intensity may impact the process of stigma and pollen recognition,and
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facilitate pollen tube growth through involvement in signal transduction and hormone synthesis between
stigma and pollen.Blueberry stigma transcriptome data analysis revealed that with the increase in
pollination intensity,the expression levels of gene CPK17and gene MIOX1were significantly
upregulated.GO enrichment analysis of differentially expressed genes identified enrichment of
functional genes associated with carbohydrate metabolic processes,inositol oxygenase activity,and cell
wall organization.Additionally,KEGG enrichment analysis highlighted significant enrichment of
pathways related to plant-pathogen interaction and inositol phosphate metabolism.
Keywords:Blueberry,Pollination,Efficiency,Transcriptome,Fruit quality
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