亚麻快速生长期细胞壁形成相关基因的表达分析

doi:10.3864/j.issn.0578-1752.2017.13.004

摘要/Abstract

摘要： 【目的】亚麻在快速生长期其韧皮纤维细胞发育在SP（the snap point）点上下端分别经历细胞伸长和次生细胞壁加厚2个不重叠时期。研究亚麻快速生长期不同组织、不同时期、不同器官中与细胞壁形成相关的β-半乳糖苷酶（LuBGALs）、纤维素合酶（LuCESAs）等家族基因的表达谱，探讨快速生长期亚麻韧皮纤维细胞细胞壁的发育模式，为改善亚麻纤维产量提供理论依据。【方法】以生长45 d的亚麻根、茎韧皮纤维、叶为材料，用透射电镜观察并测量茎韧皮纤维细胞细胞壁结构和厚度，采用实时荧光定量（qRT-PCR）方法，研究亚麻快速生长期LuBGALs和LuCESAs等细胞壁形成相关的基因在亚麻韧皮纤维不同阶段的表达特点。【结果】在SP点上部TOP端纤维细胞细胞壁薄，约110 nm；紧邻SP点茎中部的MID区（约500 nm）和茎下部的BOT区（约650 nm），细胞壁厚度明显增厚，细胞壁质地均一，没有明显的分层现象，说明SP点中部和下部韧皮纤维细胞细胞壁已经开始加厚但还未进入次生壁加厚阶段，与TOP明显不同。亚麻LuBGAL1在TOP区的表达显著低于MID区和BOT区，表明其主要参与纤维细胞细胞壁加厚过程。而LuBGAL3、LuBGAL6、LuBGAL9在TOP区表达最高，MID区次之，表明此类基因主要参与亚麻韧皮细胞伸长和细胞壁重建过程。LuBGAL5在幼嫩的TOP区表达量高，在亚麻茎较为成熟的MID区较低，说明LuBGAL5在细胞壁形成过程中起作用。其他BGALs基因的表达量均较低。在亚麻茎幼嫩的TOP区纤维细胞中，亚麻纤维素合酶基因LuCESA1、LuCESA3、LuCESA7、LuCESA8、LuCESA9和LuCESA10都检测出较高的表达量，且明显高于其在MID区和BOT区的表达。其中LuCESA3和LuCESA10在MID区的表达显著低于BOT区，其他几个CESAs基因在MID和BOT的表达并无明显差异。结合这些基因在亚麻快速生长期不同器官中的表达模式，结果说明，亚麻中6个CESA（LuCESA1、LuCESA3、LuCESA7、LuCESA8、LuCESA9和LuCESA10）主要促进亚麻韧皮纤维细胞的伸长。LuSuSy在幼茎韧皮纤维细胞中表达量高，表明亚麻茎伸长和加粗需要大量能量。LuXTH4在亚麻细胞壁发育过程中发挥作用。【结论】快速生长期亚麻茎韧皮纤维细胞细胞壁没有次生加厚过程；LuBGAL3、LuBGAL5、LuBGAL6、LuBGAL9、LuCESA1、LuCESA3、LuCESA9和LuCESA10在亚麻细胞壁细胞伸长过程中起作用；LuBGAL1主要促进亚麻细胞壁加厚过程；LuSuSy和LuXTH4在亚麻细胞壁发育中发挥作用。

关键词: 亚麻, 韧皮纤维, 细胞壁形成, 基因相对表达量

Abstract: 【Objective】 The development of flax bast cells is separated into two non-overlapping phases, cell elongation and secondary cell wall thickening phases which are splitted by the snap point at rapid-growing phase. Genes of β-glycosyl hydrolyse (LuBGALs) family and Cellulose synthase (LuCESAs) family focused on the role of cell wall formation in flax (Linum usitatissimum) fibres were analyzed in order to gain a deeper insight into the development of cell walls in rapid growth of flax.【Method】The microstructure of the cell walls at snap point (SP) of stem was studied by transmission electron microscope. LuBGALs and LuCESAs were analyzed by quantitative real-time PCR (qRT-PCR) in the bast fiber of stem, leaves and root of flax at 45 days after germination. 【Result】The width of cell walls at top of SP (TOP) was 110 nm. Though they at MID were about 500 nm and BOT about 650 nm under the SP were thicker than theirs in TOP, it was difficult to find different layers in the thicken cell wall. It showed that the cell walls above the SP were undergone cell elongation and below were thickening but not secondary cell walls thickening. The expression of LuBGAL1 at TOP was significantly lower than it’s in MID and BOT, indicating that it was mainly involved in cell wall thickening of fiber cell in the stem. While the expressions of LuBGAL3, LuBGAL6, and LuBGAL9 at TOP were the highest and the followed was MID zone, which showed that those genes mainly worked in cell wall reconstructure and bast fiber elongation. LuBGAL5 was higher at TOP and lower at MID than other genes, which demonstrated that LuBGAL5 played a role in cell wall formation. And the expression of other BGALs genes were low. LuCESA1, LuCESA3, LuCESA7, LuCESA8, LuCESA9, and LuCESA10 highly expressed in TOP zone and were significantly higher than their expression in MID and BOT. The expression of LuCESA3 and LuCESA10 in MID was lower than theirs in BOT and there was no significant difference between MID and BOT of other CESAs genes. The results suggested that these CESA genes played an important role in cell elongation of flax bast fiber combining their expression in different organs. The character of LuSuSy expression indicated that a lot of UDPG was required in cell elongation and thickening of bast fiber. LuXTH4 played a role in cell wall development of flax.【Conclusion】The cell walls of bast fiber had not processed secondary thickening in the stem at fast growth stage. LuBGAL3, LuBGAL5, LuBGAL6, LuBGAL9, LuCESA1, LuCESA3, LuCESA9, and LuCESA10 mainly promoted cell elongation of cell wall in flax. LuBGAL1 played an important role in cell wall thickening. LuSuSy and LuXTH4 also took part in the development of cell wall.

Key words: flax (Linum usitatissimum L.), bast fiber, cell wall formation, gene expression

江海霞，郭栋良，李玉环，闫文亮，杨亮杰，谢丽琼. 亚麻快速生长期细胞壁形成相关基因的表达分析[J]. 中国农业科学, 2017, 50(13): 2442-2450.

JIANG HaiXia, GUO DongLiang, LI YuHuan, YAN WenLiang, YANG LiangJie, XIE LiQiong. The Expression of Genes Related to Cell Wall Formation at Fast Growth Stage in Flax (Linum usitatissimum L.)[J]. Scientia Agricultura Sinica, 2017, 50(13): 2442-2450.

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