中文摘要：

【目的】PHT1基因家族是影响植物吸收磷营养的重要磷转运子之一。从杉木32号磷高效家系c DNA中克隆得到PHT1基因家族的1个杉木磷转运蛋白基因ClPht1;1,并对不同程度磷胁迫下ClPht1;1的时空表达进行研究,为杉木PHT1基因序列特征和功能结构的研究以及磷高效利用杉木基因型的选育奠定基础。【方法】根据PHT1基因家族序列保守性设计简并引物,以32号磷高效杉木基因型根系c DNA为模板进行扩增获得目的基因ClPht1;1的c DNA序列,使用RACE技术对目的基因进行全长克隆,并对其序列特征、同源性和编码磷转运蛋白结构进行分析。实时荧光定量PCR检测ClPht1;1在32号磷高效杉木家系根、茎、叶中的表达,检测中度缺磷胁迫下ClPht1;1在不同磷利用效率杉木4号、15号、25号、27号、28号、32号家系根系中的表达差异,以及在中度、重度缺磷胁迫下ClPht1;1在32号磷高效杉木家系根系中随时间序列的表达量变化。【结果】克隆得到1个杉木磷转运蛋白PHT1基因,命名为ClPht1;1（Gen Bank登录号：KX302006）,基因序列编码区长1 638 bp,编码545 aa的蛋白质。ClPht1;1所编码蛋白质由12个疏水的跨膜区域组成,1个疑似跨膜域。每个跨膜结构域基本由17~25个氨基酸残基组成螺旋,同时跨膜蛋白的N端和C端均位于细胞质内,保守序列位于第4个跨膜域。构成蛋白质的主要骨架是α-螺旋,无信号肽序列。ClPht1;1基因编码蛋白与日本柳杉PHT基因编码蛋白的氨基酸序列相似性达到87.0%,与胡杨、油茶、马尾松等PHT家族基因编码蛋白的氨基酸序列相似性均在75%以上。ClPht1;1基因在杉木的根、茎、叶组织中均有表达,其中在根中的表达量最高,在叶中的表达量最低。在中度缺磷胁迫下,ClPht1;1基因在杉木不同家系根部的表达量为25号〉27号〉4号〉15号〉32号〉28号。在中度和重度缺磷胁迫下,ClPht1;1基因在32号杉木家系根部的?

英文摘要：

[ Objective] PHT1 gene family is one of the important phosphorus transporters which affect the uptake of phosphorus in plants. A Cunninghamia lanceolata phosphate transporter gene of PHT1 gene family was cloned from No. 32 phosphorus efficient C. lanceolata cDNA, and the temporal and spatial expression of ClPhtl; 1 under different levels of phosphorus stress was studied to lay the foundation for the research of PHT1 gene sequence characteristics, functional structure and the selection of the C. lanceolata genotype which is efficient in phosphorus utilization. [ Method ] According to the designed degenerate primer based on the conserved sequence of PHT1 gene family, the target gene ClPhtl; 1 cDNA sequence was obtained by amplification of the root cDNA of the P-efficient family No. 32 of C. lanceolata as the template, the full length of the target gene was cloned by RACE technique, and analyzed its sequence characteristics, homology and protein structure were then analyzed. The real-time fluorescence quantitative PCR was used to detect the expression of ClPhtl; 1 in roots, stems and leaves of No. 32 C. lanceolata with high P-efficiency, differential expressions of ClPhtl; 1 in the roots of C. lanceolata families of No. 4, No. 15, No. 25, No. 27, No. 28 and No. 32 in different P-utilization etticiency under moderate P deficiency stress, and the CIPhtl; 1 expression change with time in the root system of phosphorus efficient C. lanceolata No. 32 under moderate and severe phosphorus deficiency stress. [ Result] A C. lanceolata PHTI gene was cloned and named ClPhtl; 1 （GenBank accession number KJ493165）. The coding region of the gene sequence was 1 638 bp, code 545 aa protein. ClPhtl; 1 encoded proteins were composed of 12 hydrophobic transmembrane domains, including 1 suspected transmembrane domains. Each transmembrane domain was composed of 17 -25 amino acid residues, and the N and C ends of transmembrane proteins were located in the cytoplasm, and the conserved sequences were located in the fourth transmembrane