金边红苞凤梨叶色突变体色素鉴定及类胡萝卜素合成限速基因筛选

doi:10.16420/j.issn.0513-353x.2021-0355

摘要/Abstract

摘要：

以金边红苞凤梨叶片边缘呈红色时期的嵌合体叶片和金黄色突变体叶片为材料,进行CIEL^*a^*b^*分析、色素含量测定、组织细胞观察、类胡萝卜素LC-MS/MS检测和转录组测序联合对比分析,对金黄色突变叶片的表型成因进行初步探究。CIEL^*a^*b^*分析表明,嵌合叶片中央绿色部分与边缘红色部分的颜色分别由绿、黄和红、黄颜色组成,而金黄色突变叶片为黄色中带有极微量绿色。金黄色突变叶片的颜色特征是由于其总叶绿素含量和Chl.a/b值极显著低于正常绿色部分、总花青素极显著低于边缘红色部分、类胡萝卜素和叶绿素b含量占比极显著上升形成的。金黄色突变叶片中除少量细胞含有黄绿色质体外,其他叶肉细胞为无色透明,其中共检测到12种类胡萝卜素物质,叶黄素占比高达79%,是主要呈色物质。转录—代谢联合分析表明金黄色突变叶片中将碳流转化为类胡萝卜素的八氢番茄红素合成酶基因PSY1的表达水平以及该酶下游产物八氢番茄红素、α-胡萝卜素和叶黄素等物质的含量均低于中央绿色部分,PSY1可能是金黄色突变叶片类胡萝卜素合成受到限制的关键基因。

关键词: 红苞凤梨, 金黄色突变体, 类胡萝卜素, 代谢组, 转录组

Abstract:

In order to explore how the phenotype of yellow mutants of chimeric Ananas comosus var. bracteatus formed,chimeral leaves in red stage and yellow mutants were taken in this study for comprehensive and comparative analysis using CIEL^*a^*b^* coloration,pigment determination,histocytological observation,LC-MS/MS detection and illumina RNA-seq. Results of CIEL^*a^*b^* indicated that the color of the green central part and red margin part of the chimeric leaves was consisted of green-yellow and red-yellow color,respectively. The yellow mutant showed bright yellow color with extremely slight green. The significant decrease of total chlorophyll content,Chl.a/b ratio,total anthocyanin content and the significant increase of the proportion of carotenoid and Chl.b of the yellow mutant leaf resulted in the loss of green and red color and turned to yellow. The yellow pigments were scattered in a bit of mesophyll cells of the yellow mutant leaf while the other leaf cells were transparent and colorless. Lutein accounted for 79% of the 12 detected carotenoids content which is the dominant pigment of the yellow mutant leaf. The decrease of PSY1 expression and phytoene,carotenoid,lutein content indicated that PSY1 probably was a key gene responsible for the carotenoid biosynthesis limitation in the yellow mutant of chimeric Ananas comosus var. bracteatus.

Key words: Ananas comosus var. bracteatus, yellow mutant, carotenoid, metabolome, transcriptome

中图分类号:

S668.3

周徐子鑫, 杨威, 毛美琴, 薛彦斌, 马均. 金边红苞凤梨叶色突变体色素鉴定及类胡萝卜素合成限速基因筛选[J]. 园艺学报, 2022, 49(5): 1081-1091.

ZHOU Xuzixin, YANG Wei, MAO Meiqin, XUE Yanbin, MA Jun. Identification of Pigment Components and Key Genes in Carotenoid Pathway in Mutants of Chimeric Ananas comosus var. bracteatus[J]. Acta Horticulturae Sinica, 2022, 49(5): 1081-1091.

图/表 6

图1 金边红苞凤梨嵌合体植株（A）、金黄色突变体植株（B）及不同颜色叶片和取样（C ~ E）

Fig. 1 Ananas comosus var. bracteatus yellow mutant（A）,chimeric plant（B）and different samples（C-E）

图2 金边红苞凤梨不同颜色叶片的组织细胞整体结构

Fig. 2 Tissue structure and cells of different color samples of Ananas comosus var. bracteatus

表1 金边红苞凤梨不同颜色叶片部位CIEL*a*b*指标比较

Table 1 Comparison of CIEL*a*b* indexes of Ananas comosus var. bracteatus leaf samples

样品 Sample	部位Position	L^*	a^*	b^*
金边红苞凤梨	中央绿色 Green central part	55.69 ± 1.95 c	-5.42 ± 0.25 c	17.23 ± 1.55 b
Acomosus comosus	边缘红色 Red margin part	67.62 ± 1.30 b	23.48 ± 2.89 a	17.52 ± 0.83 b
金黄色突变体Yellow mutant		72.61 ± 3.23 a	-0.30 ± 0.67 b	31.13 ± 0.54 a

图3 金边红苞凤梨不同颜色叶片部位色素含量（A）和比例（B）分析

Fig. 3 Pigment contents（A）and rates（B）in three different samples of Ananas comosus var. bracteatus

表2 金边红苞凤梨不同颜色类型叶片类胡萝卜素物质种类和相对含量的比较

Table 2 Comparison of carotenoids relative contents of three different samples of chimeric Ananas comosus var. bracteatus

分支 Branch	类胡萝卜素组分 Carotenoid	金边红苞凤梨Chimeric plant		金黄色突变体 Yellow
分支 Branch	类胡萝卜素组分 Carotenoid	中央绿色Green	边缘红色 Red	金黄色突变体 Yellow
LCYE	叶黄素Lutein	26.98 ± 0.06 a（74%）	23.37 ± 0.08 c（75%）	24.92 ± 0.13 b（79%）
	α-隐黄质α-cryptoxanthin	15.52 ± 0.08 b（< 1%）	14.20 ± 0.19 c（< 1%）	16.46 ± 0.31 a（< 1%）
	α-胡萝卜素α-carotene	20.52 ± 0.12 a（1%）	15.19 ± 0.14 b（< 1%）	16.90 ± 0.27 b（< 1%）
LCYB	β-胡萝卜素β-carotene	23.98 ± 0.03 a（10%）	19.57 ± 0.09 c（5%）	21.19 ± 0.11 b（6%）
	新黄质Neoxanthin	23.76 ± 0.04 a（9%）	18.59 ± 0.10 b（3%）	19.23 ± 0.10 b（2%）
	玉米黄质Zeaxanthin	21.21 ± 0.09 a（1%）	16.58 ± 0.09 b（1%）	16.98 ± 0.10 b（< 1%）
	紫黄质Violaxanthin	22.71 ± 0.15 a（4%）	20.77 ± 0.13 c（13%）	22.01 ± 0.11 b（11%）
	花药黄质Antheraxanthin	19.83 ± 0.14 a（1%）	15.55 ± 0.21 b（< 1%）	17.30 ± 0.11 b（< 1%）
其他Others	八氢番茄红素Phytoene	18.92 ± 0.03a（< 1%）	15.74 ± 0.12 b（< 1%）	15.65 ± 0.13 b（< 1%）
	六氢番茄红素Phytofluene	17.58 ± 0.07 b（< 1%）	17.91 ± 0.10 a（2%）	17.47 ± 0.20 b（1%）
	β-阿朴胡萝卜素醛β-apo-8'-carotenal	16.09 ± 0.04 a（< 1%）	13.47 ± 0.12 c（< 1%）	14.95 ± 0.18 b（< 1%）
	β-隐黄质β- cryptoxanthin	19.22 ± 0.04 a（< 1%）	16.52 ± 0.14 b（1%）	18.76 ± 0.48 a（1%）
	总量 Total	27.32 ± 0.05 a	23.78 ± 0.08 c	25.26 ± 0.11 b

图4 金边红苞凤梨3种颜色样品的类胡萝卜素合成的代谢—转录热图图中数字表示对应基因的FPKM值的log2对数值,* 表示组间具有差异显著性。方形色块的行数代表注释为该基因的数量。六边形中的字母表示3个样本间该物质含量的差异显著性标记。黑色无字母表示未检测到的物质。Gr代表中央绿色样品,Re代表边缘红色样品,Ye代表金黄突变样品。

Fig. 4 Transcriptome and metabolome of carotenoid biosynthesis pathway of three different samples of chimeric Ananas comosus var. bracteatus Log2 transformation represent FPKM. * represents significant difference of genes. Number of rows of color bars represent number of annotated genes. Different letters in hexagons represent significant difference among samples. The black blots represent undetectable metabolites. Gr：Green sample;Re：Red sample;Ye：Yellow mutant sample.

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