荷兰鸢尾‘玉妃’花色变异关键结构基因分析

doi:10.3864/j.issn.0578-1752.2021.12.014

Abstract

Abstract:

【Objective】 Flower color variation is of great significance for enriching color of ornamental plants, but it is difficult to clarify variation mechanism due to uncertainty of flower color variation. Dutch iris (Iris hollandica) is an important bulbous ornamental plant. In this study, the blue-purple wild type ‘Zhanchi’ and white mutant strain 'Yufei' of Dutch iris were investigated to explore molecular mechanism and difference of pigment accumulation, so as to provide a basis for mechanism of flower color variation.【Method】In this study, using inner tepals of ‘Zhanchi’ and ‘Yufei’ from Dutch Iris as materials, UHPLC-QTOF-MS method was used to determine types and contents of anthocyanins and flavonols from two varieties of Dutch Iris, and the different expression genes related to anthocyanin synthesis were screened by transcriptome sequencing. Using different flowers in developmental stages of two varieties as materials, the different expression genes were verified by qRT-PCR.【Result】Results of metabolomics analysis revealed that delphinidin, cyanidin and its derivatives were accumulated in blue-purple flowers, which were almost no accumulation in white flowers, while the flavonol contents were increased in white ‘Yufei’. Results of RNA-seq analysis revealed that a total of 46 485 unigenes were obtained, and 27 073 unigenes of them were functionally annotated by public databases, accounting for 41.85% of the total. And 701 differentially expressed genes were obtained, 485 genes of which were up-regulated and 216 genes were down-regulated in white ‘Yufei’. Two dihydroflavonol-4-reductase genes and one flavonol synthase gene involving in anthocyanin biosynthetic pathway had different expression, named IhDFR1, IhDFR2 and IhFLS1. Down-regulated expression of IhDFR1 and IhDFR2 as well as up-regulated expression of IhFLS1 in white ‘Yufei’ led to significant decrease of anthocyanins and accumulation of flavonols, which caused metabolic flow from anthocyanin to flavonol. The qRT-PCR results of three genes showed that expression levels of IhDFR1 and IhDFR2 increased in blue-purple flowers during flower developmental stage, but low expression in white flowers, and expression level of IhFLS1 increased in white flowers during flower developmental stage, but low expression in blue-purple flowers, which was consistent with RNA-seq results.【Conclusion】Low expression of IhDFR1 and IhDFR2 as well as high expression of IhFLS1 in white ‘Yufei’ blocked accumulation of anthocyanins, and some of metabolic flow changed from anthocyanins to flavonols, resulting in the change of flower color from blue violet to white.

Key words: Iris hollandica, anthocyanin, flavonol, transcriptome analyses, dihydroflavonol-4-reductase gene, flavonol synthase gene

LIN Bing,CHEN YiQuan,ZHONG HuaiQin,YE XiuXian,FAN RongHui. Analysis of Key Genes About Flower Color Variation in Iris hollandica[J].Scientia Agricultura Sinica, 2021, 54(12): 2644-2652.

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Figures/Tables 6

Table 1

Fig. 1

Fig. 2

Table 2

Identification of anthocyanin and flavonol in Iris hollandica flower"

化合物种类 Species of compounds	Compound 化合物名称	含量 Content (μg·g^-1)
化合物种类 Species of compounds	Compound 化合物名称	展翅 Zhanchi	玉妃 Yufei
花青素苷Anthocyanidin	矢车菊素-3-(2G-木糖基芸香糖苷) Cyanidin -3-(2G-xylosylrutinoside)	25.22±1.66	1.34±0.05
	飞燕草素-3-[6-(4-咖啡酰基木糖基)葡萄糖苷]-5-葡萄糖苷 Delphinidin-3-[6-(4-(caffeoylrhamnosyl) glucoside]-5-glucoside	61.26±1.33	1.36±0.08
	矢车菊素-3-芸香糖苷 Cyanidin -3-rutinoside	144.42±6.46	16.90±0.63
	6-羟基矢车菊素-3-葡萄糖苷 6-Hydroxycyanidin -3-glucoside	5.88±0.19	0.56±0.01
	飞燕草素-3-(6-p-酰基葡萄糖苷)-5-(6-丙二酰基-4-鼠李糖基葡萄糖苷) Delphinidin-3-(6-p-coumaroylglucoside)-5-[6-(malonyl)-4-(rhamnosyl) glucoside)]	209.85±3.12	1.71±0.05
	飞燕草素-3-葡萄糖苷 Delphinidin-3-glucoside	4.37±0.24	0.08±0.01
	飞燕草素-3-芸香糖苷 Delphinidin 3-rutinoside	30.64±0.38	16.98±0.48
	矮牵牛素-3-(6-鼠李糖基-2-木糖基葡萄糖苷) Petunidin-3-[6-(rhamnosyl)-2-(xylosyl) glucoside]	61.86±0.59	8.29±0.20
	总计 Total	543.5	47.22
黄酮醇 Flavonol	山奈酚-3-O-半乳糖苷 Kaempferol-3-O-galactoside	73.37±2.13	69.21±1.69
	山奈酚-7-O-葡萄糖苷 Kaempferol-7-O-glucoside	62.36±2.61	56.64±0.85
	槲皮素-3-O-葡萄糖苷 Quercetin-3-O-glucoside	21.45±0.32	12.81±0.24
	鼠李素-3-O-葡萄糖苷 Rhamnetin-3-O-Glucoside	40.01±0.51	22.95±0.32
	异鼠李素-3-O-葡萄糖苷 Isorhamnetin-3-O-Glucoside	37.02±1.20	24.14±0.67
	苜蓿素-4'-甲基醚-3'-O-葡萄糖苷 Tricin-4'-methylether-3'-O-glucoside	39.31±0.45	23.55±0.33
	杨梅素-3-O-葡萄糖苷 Myricetin-3-O-glucoside	3.29±0.08	145.22±3.74
	杨梅素-3-O-半乳糖苷 Myricetin-3-O-galactoside	4.04±0.12	58.65±2.52
	槲皮素-3-O-(6''-丙二酰)半乳糖苷 Quercetin-3-O-(6''-malonyl)galactoside	4.99±0.12	6.27±0.32
	鼠李素-3-O-芸香糖苷 Rhamnetin-3-O-Rutinoside	4.36±0.21	15.05±1.23
	3,5,7,4'-四羟基-8-甲氧基黄酮-3-O-葡萄糖苷-7-O-鼠李糖苷 Sexangularetin-3-O-glucoside-7-O-rhamnoside	4.48±0.09	14.39±0.41
	羟基山奈酚-3,6-O-二葡萄糖苷 6-Hydroxykaempferol-3,6-O-Diglucoside	4.89±0.25	1.21±0.06
	槲皮素-3-O-芸香糖苷-7-O-鼠李糖苷 Quercetin-3-O-rutinoside-7-O-rhamnoside	28.55±1.01	3.75±0.31
	槲皮素-7-O-芸香糖苷-4'-O-葡萄糖苷 Quercetin-7-O-rutinoside-4'-O-glucoside	5.04±0.06	0.02±0.01
	总计 Total	333.15	453.81

Table 2

Table 3

Fig. 3

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基因 Gene	正向引物（5′-3′）Forward primer sequence	反向引物（5′-3′）Reverse primer sequence
IhDFR1	GAGGTGGTCGCAGGATGCACT	CTCCGTCTGCTGATGTTCTTT
IhDFR2	AAGGCGGTCGCAGGATGCACC	TGATGAAAGGACCGACGACT
IhFLS1	GTTGGAGTGATGGACGGGATG	GGGACAGGGAGGGTAGTAGTTGA
Ihactin	ACGGAAATTGTATGTGGGT	CAGATGCGAAAGATGTGAG

样品 Sample	Clean read 数量 Number of clean reads	GC含量 GC content (%)	%≥Q30
展翅 Zhanchi	45850456	50.29	87.76%
玉妃 Yufei	48903721	50.30	87.08%

Analysis of Key Genes About Flower Color Variation in Iris hollandica

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