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长牡蛎温度应激响应相关基因的初步研究
竺奇慧1,2
Subtype博士
Thesis Advisor张国范 ; 李莉
2015-11
Degree Grantor中国科学院大学
Place of Conferral北京
Degree Discipline海洋生物学
Keyword牡蛎 高温 低温 基因表达 Chip-seq 热激蛋白
Abstract长牡蛎(Crassostrea gigas)是一种典型的在潮间带营固着生活的物种,它
的生活环境决定了它要遭受来自环境的强烈应激。因为需要长时间暴露在空气中,
牡蛎对温度变化有一个很广的适应范围。但是到目前为止,对于长牡蛎的广温适
应性机制,无论是细胞层面还是分子层面,都鲜有深入的研究。
在本研究中,我们利用本实验前期所做的长牡蛎基因组数据以及温度应激转
录组数据,进一步分析了长牡蛎在高温以及低温下的基因表达,筛选出了33 个
温度应激相关基因,其中主要包括热激蛋白基因以及凋亡相关基因等;另一方面,
我们选取了10 个应激相关的基因家族,在11 个不同物种之间分析了它们的基因
数量以及序列,着重分析了长牡蛎基因组中热激蛋白70 家族基因。
对于筛选出的33 个温度应激相关基因,我们分别设计了长牡蛎高温(35℃)
和低温(5℃)应激实验,用荧光定量PCR 方法研究了这些基因的表达模式,结
果发现:(1)在高温应激实验中,受高温诱导表达的基因在应激的0.5 h 表达量
就显著上升,但是它们的表达峰值却是在应激恢复期出现;(2)这些基因在短时
间低温应激中表达量变化较小,但是在长时间低温应激的3 d 左右时出现最高表
达量;(3)在经历2 天的应激恢复期后,再次对长牡蛎做相同的高温应激时,这
些受高温诱导基因的表达量变化却很小,而且此时牡蛎的死亡率也比第一次应激
时要低很多。这些结果说明牡蛎对高温应激具有可塑性,且这种可塑性与温度应
激相关基因的表达有关。我们也进一步确定了热激蛋白基因和凋亡相关蛋白在长
牡蛎抗应激系统中的重要作用。对于10 个应激相关的基因家族在11 个不同物种
之间分析结果表明,这些基因家族存在谱系特异的扩张,这说明基因家族扩张与
跟物种的适应性相关,即暗示了物种的适应性进化。综合上述结果,由基因表达
而表现出的可塑性以及适应性进化两方面原因可能在长牡蛎抗应激系统的发展
中起到重要的作用。
已有文献报道认为,热激因子HSF1 可能是调控HSP 在受应激时表达的重
要转录因子之一。我们通过ChIP-seq 方法对HSF1 所结合的DNA 序列进行了分
析,结果发现5 个HSP 蛋白与HSF1 存在相互作用,除热激蛋白外,HSF1 可能
还调控其他基因,它可能在长牡蛎抵抗多种不同应激的过程中起到重要作用。
Other AbstractAs a characteristic sessile inhabitant of the intertidal zone, the Pacific oyster
Crassostrea gigas occupies one of the most physically stressful environments on earth.
With high exposure to terrestrial conditions, oysters must tolerate broad fluctuations
in temperature range. However, oysters’ cellular and molecular responses to
temperature stresses have not been fully characterized.
In this study, we analyzed oyster transcriptome data under high and low
temperatures. We also identified over 30 key gene candidates, which encoded stress
proteins such as heat shock proteins and apoptosis-associated genes that responded to
temperature stresses. The characterization of the expression patterns of these genes
under short-term cold and hot environments (5°C and 35°C) and long-term cold
environments (5°C) were detected by quantitative real-time PCR. The expression of
most temperature-responsive genes reached peaks during the recovery stage after heat
stress, and these genes were most perturbed around 3 days and responded little to
short-term cold stress. In addition, oysters in the second heat challenge after 2 days of
recovery showed a milder expression of the temperature-responsive genes and lower
mortality rate, which indicated the existence of plasticity in the oyster’s response to
heat stress. We confirmed that homeostatic flexibility and anti-apoptosis might be
crucial centers of temperature stress responses in oysters. In addition, we analyzed
stress protein families in 11 different species and found that the linage-specific
expansion of stress genes might be implicated in adaptive evolution. The results
indicated that both plasticity and evolution play an important role in the
stress-response adaptation of oysters.
It has been proposed that the transcription of HSP genes is induced by the release
of transcription factor HSF1. We analyzed the DNA sequence that HSF1 bound to by
ChIP method and found that besides some HSPs, HSF1 might regulate many genes
that involved in various cellular processes.
Subject Area分子生物学 ; 水产生物学 ; 水产学基础学科其他学科
Language中文
Document Type学位论文
Identifierhttp://ir.qdio.ac.cn/handle/337002/95165
Collection海洋生物技术研发中心
Affiliation1.中国科学院海洋研究所
2.中国科学院大学
Recommended Citation
GB/T 7714
竺奇慧. 长牡蛎温度应激响应相关基因的初步研究[D]. 北京. 中国科学院大学,2015.
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