苹果纯合基因型新种质的性状鉴定与培养

doi:10.3864/j.issn.0578-1752.2018.10.015

摘要/Abstract

摘要：

目的苹果花药培养获得的纯合基因型株系来自不同的花粉粒,因此不同株系的植物学特征、生物学特性,以及诱导培养、生根的条件等都存在差异。本研究对不同个株系试管苗的倍性、表型特征、生根培养条件进行观察分析,选出苹果纯合基因型株系适宜的生根条件及综合性状较好的株系,加强苹果种质资源创新。方法采用花药培养获得的‘嘎啦’‘富士’‘红星’纯合基因型株系,利用流式细胞仪分析每个株系倍性,探索组培苗的生根培养条件,观察每个株系的根系、叶片形态特征。结果经苹果花药培养获得32个纯合基因型株系,其中单倍体1株、三倍体1株、四倍体3株、二倍体27株,‘红星’苹果纯合基因型株系的倍性分化率最大,为28.57%。IBA浓度影响再生株系的生根率、根长及根数,苹果纯合基因型株系的最适宜生根浓度为IBA 2—3 mg·L^-1。各个株系的叶数、根长、根数、株高、叶形指数（叶长度/叶宽度）、叶柄有一定差异。‘红星’纯合基因型DH0-1、DH0-3、DH0-4、DH0-7株系,‘富士’纯合基因型DH1-3株系,‘嘎啦’纯合基因型DH2-2、DH2-4、DH2-12、DH2-20株系综合性状较好（植株高、根系长、叶数和根数多）。‘红星’纯合基因型株系的移栽成活率最高,为28.57%。结论苹果纯合基因型株系的主要倍性为二倍体。与二倍体株系相比,单倍体株系叶基窄、叶柄细、叶色和叶缘锯齿较浅,而多倍体株系表现为叶基更阔、叶柄较粗、叶色和叶缘锯齿变深。

关键词: 苹果花药培养, 倍性分析, 纯合基因型, 生根激素, 性状分析

Abstract:

【Objective】 Homozygous genotype lines were obtained by anther culture of different apple pollen grains, so their botanical characteristics, biological characteristics, and their induced culture, root conditions were different. In this study, the ploidy, characteristics and root conditions of each line were observed and analyzed. Appropriate rooting conditions and good combinations of lines for selecting apple homozygous genotypes are of great significance to the innovation of apple germplasm resources.【Method】 The homozygous genotype lines of ‘Gala’, ‘Fuji’ and ‘Red Star’ were obtained by apple anther culture. The ploidy level of homozygous genotype lines was studied using flow cytometry, root and leaf morphology and root conditions of each line were observed and recorded. 【Result】 A total of 32 homozygous lines were obtained by apple anther culture. The results of flow cytometry showed that there were 1 haploid, 1 triploids, 3 tetraploids and 27 diploids. The ploidy rate of 'Red Star' homozygous genotype was the highest, about 28.57%. The root rate, root length and root number of homozygous lines were affected by IBA concentration of rooting medium. When the concentration of IBA was 2-3 mg·L^-1, the root features of apple homozygous lines were the best. The leaf number, root length, root number, plant height, leaf length/width and petiole length of each line were different. ‘Red Star’ homozygous genotype lines (DH2-1, DH0-3, DH0-4, DH0-7), ‘Fuji’ homozygous genotype line (DH1-3) and ‘Gala’ homozygous genotype lines (DH2-2, DH2-4, DH2- 12, DH2-20) have a good traits (plants are taller, roots are longer, and the number of roots and leaves are more). The survival rate of ‘Red Star’ homozygous genotype was the highest, about 28.57%. 【Conclusion】 The main ploidy of apple homozygous genotype was diploid. Compared with the diploid lines, the haploid lines had a narrow leaf base, finer petiole, the leaf color and the leaf margin were shallow, while the polyploid lines showed wider leaf base, thicker petioles, leaf color and leaf edge serrated deep.

Key words: apple anther culture, ploidy analysis, homogeneous genotype, root hormone, trait analysis

石江鹏, 张春芬, 邓舒, 侯丽媛, 肖蓉, 李芙蓉, 董艳辉, 聂园军, 王亦学, 曹秋芬. 苹果纯合基因型新种质的性状鉴定与培养[J]. 中国农业科学, 2018, 51(10): 1960-1971.

JiangPeng SHI, ChunFen ZHANG, Shu DENG, LiYuan HOU, Rong XIAO, FuRong LI, YanHui DONG, YuanJun NIE, YiXue WANG, QiuFen CAO. Morphological Identification and Cultivation of New Germplasm of Apple Homozygous Genotypes[J]. Scientia Agricultura Sinica, 2018, 51(10): 1960-1971.

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https://www.chinaagrisci.com/CN/Y2018/V51/I10/1960

图/表 12

表1

图1

表2

图2

表3

表4

不同IBA浓度对‘嘎啦’苹果纯合基因型株系根系形态的影响"

根系形态 Root morphology	培养基 Medium	株系 Line
根系形态 Root morphology	培养基 Medium	DH2-1	DH2-2	DH2-3	DH2-4	DH2-5	DH2-6	DH2-7	DH2-8	DH2-9	DH2-10
生根率 Root rate (%)	T1	6.67	100	20	_	6.67	80	100	53.33	66.67	20
	T2	60	100	66.67	100	_	100	100	73.33	86.67	60
	T3	13.33	90	80	86.67	13.33	100	100	73.33	86.67	86.67
根长 Root length (cm.plant-1)	T1	4.65±0.97a	1.70±0.67b	2.93±0.49b	_	5.9a	4.27±0.88b	0.57±0.39b	32.60±1.05a	9.50±1.25a	7.53±1.31b
	T2	6.43±1.47a	1.63±0.78b	1.87±1.19b	29.43±1.80a	_	8.90±1.07a	2.50±1.02a	30.00±1.10a	8.93±1.25a	8.23±1.76b
	T3	1.5a	5.77±1.17a	13.17±1.20a	14.83±1.40b	5.85±1.18a	9.40±1.15a	3.67±0.71a	31.33±1.28a	10.93±1.54a	22.10±1.31a
根数 Root number	T1	1.50±0.89b	24.67±1.65b	3.67±1.15a	_	1a	9.33±0.83c	16c	9.00±1.20b	5.67±1.45a	1.33±0.83b
	T2	17.00±1.66a	39.67±1.67ab	1.67±0.83a	4.00±1.38a	_	42.33±1.82a	29.33±1.83b	21.67±1.61a	9.67±1.45a	5.00±1.20b
	T3	1b	51.67±2.42a	2.33±0.83a	6.00±1.38a	1a	34.67±1.28b	43.67±1.73a	8.00±1.59b	12.00±1.74a	22.33±1.86a
根系形态 Root morphology	培养基 Medium	株系 Line
根系形态 Root morphology	培养基 Medium	DH2-11	DH2-12	DH2-13	DH2-14	DH2-15	DH2-16	DH2-17	DH2-18	DH2-19	DH2-20
生根率 Root rate(%)	T1	66.67	20	80	6.67	53.33	_	6.67	93.33	73.33	6.67
	T2	100	100	100	73.33	66.67	20	53.33	100	73.33	86.67
	T3	93.33	53.33	100	60	53.33	_	13.33	100	93.33	66.67
根长 Root length (cm.plant-1)	T1	14.5±1.62b	12.7±1.26b	1.33±0.69c	4.8b	14.27±1.46a	_	4.5b	7.60±1.30c	13.43±1.54a	13.8a
	T2	20.7±1.40a	35.27±1.78a	3.83±1.18b	17.33±1.63a	10.23±1.25a	3.43±1.02	11.77±1.40a	16.10±1.72b	14.43±1.38a	18.40±1.57a
	T3	20.7±1.40a	23.2±1.90b	6.27±0.96a	6.40±1.36b	9.80±1.26a	_	3.20±1.46b	23.83±1.42a	11.07±1.33a	14.77±1.48a
根数 Root number	T1	3.00±1.20b	2.33±0.83b	11.67±0.83b	1a	13.67±1.36a	_	1b	5.00±1.00b	3.33±1.15b	27c
	T2	10.33±1.36a	11.33±1.15a	18.67±1.59ab	6.33±1.36a	7.00±1.63b	1.33±0.83	2.67±0.83a	8.33±1.67ab	3.67±1.15b	86.67±2.05a
	T3	5.33±1.59ab	3.00±1.20b	22.33±1.71a	5.33±1.48a	5.00±1.26b	_	1.50±0.89ab	14.33±1.61a	16.00±1.53a	59.67±2.50b

表4

图3

表5

表6

表7

‘嘎啦’苹果纯合基因型株系性状"

株系 Line	叶数 Leave number	根数 Root number	根长 Root length(cm)	株高 Plant height	叶柄长 Petiole length (cm)	叶形指数 Leaf shape index
DH2-1	13.67±1.48gh	17.00±1.66ef	6.43±1.47ghi	1.70±0.46g	0.70±0.46ef	2.15±0.60bc
DH2-2	20.00±1.00defg	41.67±1.52b	6.33±1.01ghi	5.57±1.01a	1.27±0.59ab	2.29±0.57b
DH2-3	32.33±1.83abc	3.67±1.15hij	13.17±1.20f	3.17±0.71d	1.0±0.67bcde	3.07±0.68a
DH2-4	36.67±2.30a	5.00±1.00ghij	27.17±2.08bc	2.97±0.86de	1.33±0.39a	1.74±0.34defg
DH2-5	11.33±1.36h	1j	5.87±1.05ghi	1.70±0.70g	0.57±0.39f	1.69±0.75efg
DH2-6	21.33±1.28defg	35.67±1.15c	9.97±0.98fgh	3.20±0.71d	0.80±0.64cdef	2.21±0.45b
DH2-7	18.67±1.28defgh	43.67±1.72b	35.27±1.78a	5.10±0.58ab	1.10±0.46abc	2.07±0.51bcd
DH2-8	20.67±1.93defg	21.67±1.61de	3.67±0.71i	1.67±0.53g	0.80±0.64cdef	1.92±0.68bcdef
DH2-9	19.33±1.45defgh	8.00±1.26ghi	11.67±1.40fg	4.10±0.46c	0.83±0.53cdef	1.63±0.56fg
DH2-10	20.00±1.53defg	5.00±1.20ghij	22.10±1.31cd	2.20±0.64fg	0.83±0.49cdef	1.92±0.54bcdef
DH2-11	17.33±1.05efgh	10.00±1.44gh	12.53±1.31f	2.90±0.64de	0.90±0.46cde	1.40±0.45g
DH2-12	27.00±1.53bcd	11.33±1.15fg	19.63±1.36de	2.90±0.64de	0.73±0.53def	1.58±0.47fg
DH2-13	21.67±1.05defg	24.67±1.28d	4.93±1.47hi	1.77±0.53g	0.70±0.46ef	2.21±0.37b
DH2-14	25.00±1.38cde	6.33±1.36ghij	9.17±1.46fghi	2.53±0.53def	0.83±0.63cdef	2.12±0.59bc
DH2-15	20.67±1.36defg	4.00±1.00hij	14.89±1.28ef	2.47±0.39ef	0.93±0.39cde	1.80±0.55cdef
DH2-16	15.33±1.36fgh	1.33±0.83j	3.43±1.02i	3.17±0.59d	1.03±0.39bcd	2.29±0.59b
DH2-17	21.00±1.26defg	2.67±0.83ij	11.77±1.40fg	2.40±0.56ef	0.83±0.49cdef	2.11±0.39bc
DH2-18	34.33±1.78ab	11.00±1.20fg	9.87±1.03fgh	3.07±0.67de	0.80±0.58cdef	2.07±0.49bcd
DH2-19	25.00±1.38cde	11.33±1.68fg	11.17±1.31fg	3.17±0.71d	0.90±0.46cde	2.00±0.44bcde
DH2-20	23.67±1.61def	85.67±2.16a	30.00±1.10b	4.60±0.67bc	1.03±0.53bcd	2.21±0.53b

表7

图4

图5

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红星株系 Red star strain	倍性 Ploidy	富士株系 Fuji strain	倍性 Ploidy	嘎啦株系 Gala strains	倍性 Ploidy	嘎啦株系 Gala strains	倍性 Ploidy	嘎啦株系 Gala strains	倍性 Ploidy
DH0-1	2n	DH1-1	2n	DH2-1	2n	DH2-8	2n	DH2-15	4n
DH0-2	2n	DH1-2	4n	DH2-2	2n	DH2-9	2n	DH2-16	2n
DH0-3	2n	DH1-3	2n	DH2-3	1n	DH2-10	2n	DH2-17	2n
DH0-4	2n	DH1-4	2n	DH2-4	2n	DH2-11	2n	DH2-18	2n
DH0-5	4n	DH1-5	2n	DH2-5	2n	DH2-12	2n	DH2-19	2n
DH0-6	2n			DH2-6	2n	DH2-13	2n	DH2-20	2n
DH0-7	3n			DH2-7	2n	DH2-14	2n

根系形态 Root morphology	培养基 Medium	株系 Line
根系形态 Root morphology	培养基 Medium	DH0-1	DH0-2	DH0-3	DH0-4	DH0-5	DH0-6	DH0-7
生根率 Root rate (%)	T1	26.67	6.67	70	46.67	33.33	46.67	33.33
	T2	53.33	86.67	100	93.33	86.67	66.67	100
	T3	73.33	73.33	93.33	93.33	100	53.33	66.67
根长 Root length (cm/plant)	T1	5.70±0.81a	4.80b	18.13±1.53a	3.70±1.16a	3.50±0.58b	3.50±0.79a	3.90±0.56a
	T2	3.87±1.13a	6.63±1.19a	20.83±1.66a	3.73±0.77a	4.63±0.91b	3.17±0.77a	3.93±0.67a
	T3	4.60±0.71a	4.50±0.67b	20.13±1.51a	4.07±1.13a	8.33±1.17a	3.13±0.90a	4.27±0.95a
根数 Root number	T1	7.00±1.26b	3.00b	5.00±1.00b	2.33±1.05c	3.33±0.83b	3.33±1.32b	1.00c
	T2	4.67±1.32b	5.00±1.00a	10.00±1.26a	15.33±1.28a	5.67±1.05a	9.00±1.00a	22.33±1.59a
	T3	17.67±1.56a	1.33±0.83c	7.00±1.26ab	9.33±0.83b	6.33±1.15a	8.67±0.83a	12.33±1.45b

株系 Line	叶数 Leave number	根数 Root number	根长 Root length (cm)	株高 Plant height (cm)	叶柄长 Petiole length (cm)	叶形指数 Leaf shape index
DH0-1	27.00±1.00a	17.67±1.56a	4.27±0.91a	3.70±0.91ab	0.83±0.49a	1.66±0.52b
DH0-2	12.33±1.45b	4.33±1.28c	5.73±1.15b	2.37±0.80c	0.5±0.46c	2.07±0.49a
DH0-3	22.33±1.28a	10.67±1.28b	18.17±1.89a	4.13±0.80a	0.80±0.46a	1.60±0.51b
DH0-4	26.67±1.71a	18.67±1.56a	3.17±0.85b	3.47±0.71abc	0.87±0.39a	1.74±0.54b
DH0-5	15.00±1.00b	8.00±1.00bc	3.53±0.91b	2.57±0.86bc	0.63±0.39bc	1.71±0.37b
DH0-6	26.00±1.84a	11.00±1.38b	2.87±0.49b	4.57±0.49a	0.7±0.46b	1.68±0.59b
DH0-7	22.67±1.28a	22.33±1.59a	2.67±1.08b	3.67±1.01ab	0.77±0.39a	1.88±0.67ab

株系 Line	叶数 Leave number	根数 Root number	根长 Root length(cm)	株高 Plant height (cm)	叶柄长 Petiole length (cm)	叶形指数 Leaf shape index
DH1-1	15.33±1.32b	17.67±1.56b	7.93±0.87b	1.97±0.67d	0.83±0.39bc	1.90±0.63ab
DH1-2	16.00±1.38b	12.33±1.28c	22.27±1.63a	2.23±0.63cd	1.53±0.77a	1.61±0.41b
DH1-3	17.67±1.15ab	26.00±1.53a	19.00±1.56a	3.07±0.69ab	1.17±0.53ba	1.58±0.73b
DH1-4	22.67±1.28a	4.00±1.38d	4.47±1.32b	2.57±0.59bc	0.87±0.39bc	2.12±0.55a
DH1-5	22.33±1.65a	1.33±0.83d	4.43±1.55b	3.33±0.72a	0.70±0.58c	1.52±0.57b