Identification of 21 New Camellia Hybrid Varieties by Fluorescence-labelled Simple Sequence Repeat Markers

doi:10.11983/CBB18019

Abstract

Abstract: In this study, 12 highly polymorphic simple sequence repeat (SSR) loci were selected from Camellia azalea transcriptome data and previous studies, and the genotypes of 21 new Camellia hybrid varieties were analyzed by fluorescence capillary electrophoresis. Repeatable and distinct amplification products could be obtained at 12 SSR loci from all samples, and 21 new varieties could be identified by the specific genotype of 12 SSR loci. Significantly, for the 21 new varieties, the number of discrepant loci was 2 to 10 among varieties from the same parental combination but 5 to 12 among varieties from different combinations. Hence, each of the 21 varieties was marked by a unique group of molecular markers, which can be used to identify these varieties accurately. These markers are very useful for identifying and protecting Camellia varieties, especially for those propagated that are produced by grafting or cutting.

Key words: Camellia, fluorescence labelled SSR, new variety protection, variety identification

Naiqi Tao,Bin Zhang,Xinkai Liu,Heda Zhou,Naisheng Zhong,Danfeng Yan,Min Zhang,Jiyin Gao,Wenju Zhang. Identification of 21 New Camellia Hybrid Varieties by Fluorescence-labelled Simple Sequence Repeat Markers[J]. Chinese Bulletin of Botany, 2019, 54(1): 37-45.

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URL: https://www.chinbullbotany.com/EN/10.11983/CBB18019

https://www.chinbullbotany.com/EN/Y2019/V54/I1/37

Figures/Tables 7

References 38

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Cross combination No.	New variety No.	Parental types of cross combinations
ZH-01	SJ-01	Var. B (♀)×Var. A (♂)
	SJ-02	Var. B (♀)×Var. A (♂)
	SJ-03	Var. B (♀)×Var. A (♂)
	SJ-04	Var. B (♀)×Var. A (♂)
ZH-02	SJ-05	Var. C (♀)×Var. A (♂)
	SJ-06	Var. C (♀)×Var. A (♂)
	SJ-07	Var. C (♀)×Var. A (♂)
	SJ-08	Var. C (♀)×Var. A (♂)
ZH-03	SJ-09	Var. D (♀)×Var. A (♂)
	SJ-10	Var. D (♀)×Var. A (♂)
	SJ-11	Var. D (♀)×Var. A (♂)
ZH-04	SJ-12	Var. E (♀)×Var. A (♂)
ZH-05	SJ-13	Var. F (♀)×Var. A (♂)
ZH-06	SJ-14	Var. A (♀)×Var. G (♂)
	SJ-15	Var. A (♀)×Var. G (♂)
	SJ-16	Var. A (♀)×Var. G (♂)
ZH-07	SJ-17	Var. A (♀)×Var. H (♂)
ZH-07	SJ-18	Var. A (♀)×Var. H (♂)
ZH-08	SJ-19	Var. A (♀)×Var. I (♂)
ZH-09	SJ-20	Var. A (♀)×Var. J (♂)
ZH-10	SJ-21	Var. A (♀)×Var. K (♂)

Cross combination No.	New variety No.	Parental types of cross combinations
ZH-01	SJ-01	Var. B (♀)×Var. A (♂)
	SJ-02	Var. B (♀)×Var. A (♂)
	SJ-03	Var. B (♀)×Var. A (♂)
	SJ-04	Var. B (♀)×Var. A (♂)
ZH-02	SJ-05	Var. C (♀)×Var. A (♂)
	SJ-06	Var. C (♀)×Var. A (♂)
	SJ-07	Var. C (♀)×Var. A (♂)
	SJ-08	Var. C (♀)×Var. A (♂)
ZH-03	SJ-09	Var. D (♀)×Var. A (♂)
	SJ-10	Var. D (♀)×Var. A (♂)
	SJ-11	Var. D (♀)×Var. A (♂)
ZH-04	SJ-12	Var. E (♀)×Var. A (♂)
ZH-05	SJ-13	Var. F (♀)×Var. A (♂)
ZH-06	SJ-14	Var. A (♀)×Var. G (♂)
	SJ-15	Var. A (♀)×Var. G (♂)
	SJ-16	Var. A (♀)×Var. G (♂)
ZH-07	SJ-17	Var. A (♀)×Var. H (♂)
ZH-07	SJ-18	Var. A (♀)×Var. H (♂)
ZH-08	SJ-19	Var. A (♀)×Var. I (♂)
ZH-09	SJ-20	Var. A (♀)×Var. J (♂)
ZH-10	SJ-21	Var. A (♀)×Var. K (♂)

No.	Primer sequences (5′-3′)	Repeat motifs	Ta (°C)	References
478	F: CAACACCACCAACAAGA	(AAAGG)4	53	Liu et al., 2008
478	R: GATATGAGATCCGTCCC	(AAAGG)4	53	Liu et al., 2008
SSR2	F: TATTGCCTACGACCATTTCCA	(GA)14	56	Kaundun and Matsumoto, 2002
SSR2	R: TTTGAGTTCGTTGCCTTCTCT	(GA)14	56	Kaundun and Matsumoto, 2002
CamsinM11	F: GCATCATTCCACCACTCACC	(CA)12	60	Freeman et al., 2004
CamsinM11	R: GTCATCAAACCAGTGGCTCA	(CA)12	60	Freeman et al., 2004
CamSSR01	F: CCAACAAGAATCAGGAAGAG	(AAT)6	54	In this study
CamSSR01	R: ATCCAACGGTGGTAGACGAG	(AAT)6	54	In this study
CamSSR02	F: AGTTCCGCCTCCAGTTTGAC	(ACG)7	54	In this study
CamSSR02	R: GGACCGAGAGGTAACAGTGG	(ACG)7	54	In this study
CamSSR03	F: GCCACTACCCTCTTTACACC	(CAC)7	55	In this study
CamSSR03	R: TTCTCTTCCTCTTTCTTCCC	(CAC)7	55	In this study
CamSSR04	F: ATGTGTTGAGTAGCGAGCGT	(AT)10	56	In this study
CamSSR04	R: TTGTCCATCTTTATGTAGGG	(AT)10	56	In this study
CamSSR05	F: GCAAACACCAACTGATTACC	(TA)10	56	In this study
CamSSR05	R: TTCCATACAACTCAACCAAA	(TA)10	56	In this study
CamSSR06	F: GGTTTGGAAAAAGGACACGC	(GCC)7	58	In this study
CamSSR06	R: AATCTGCCTCTGGTAGTCCG	(GCC)7	58	In this study
CamSSR07	F: TCTCATCCCCATCTTTATCC	(TCC)7	58	In this study
CamSSR07	R: GTTCCCTGCTGCTGTTGTTA	(TCC)7	58	In this study
CamSSR08	F: TCACCAGTCACTTTCCCTCC	(AC)10	58	In this study
CamSSR08	R: CCACCAAAAGGCACAATACC	(AC)10	58	In this study
CamSSR09	F: CATCATCCATCAAACCGTCC	(AT)10	58	In this study
CamSSR09	R: GAAGGCACATTGGTTCTGGG	(AT)10	58	In this study

No.	Primer sequences (5′-3′)	Repeat motifs	Ta (°C)	References
478	F: CAACACCACCAACAAGA	(AAAGG)4	53	Liu et al., 2008
478	R: GATATGAGATCCGTCCC	(AAAGG)4	53	Liu et al., 2008
SSR2	F: TATTGCCTACGACCATTTCCA	(GA)14	56	Kaundun and Matsumoto, 2002
SSR2	R: TTTGAGTTCGTTGCCTTCTCT	(GA)14	56	Kaundun and Matsumoto, 2002
CamsinM11	F: GCATCATTCCACCACTCACC	(CA)12	60	Freeman et al., 2004
CamsinM11	R: GTCATCAAACCAGTGGCTCA	(CA)12	60	Freeman et al., 2004
CamSSR01	F: CCAACAAGAATCAGGAAGAG	(AAT)6	54	In this study
CamSSR01	R: ATCCAACGGTGGTAGACGAG	(AAT)6	54	In this study
CamSSR02	F: AGTTCCGCCTCCAGTTTGAC	(ACG)7	54	In this study
CamSSR02	R: GGACCGAGAGGTAACAGTGG	(ACG)7	54	In this study
CamSSR03	F: GCCACTACCCTCTTTACACC	(CAC)7	55	In this study
CamSSR03	R: TTCTCTTCCTCTTTCTTCCC	(CAC)7	55	In this study
CamSSR04	F: ATGTGTTGAGTAGCGAGCGT	(AT)10	56	In this study
CamSSR04	R: TTGTCCATCTTTATGTAGGG	(AT)10	56	In this study
CamSSR05	F: GCAAACACCAACTGATTACC	(TA)10	56	In this study
CamSSR05	R: TTCCATACAACTCAACCAAA	(TA)10	56	In this study
CamSSR06	F: GGTTTGGAAAAAGGACACGC	(GCC)7	58	In this study
CamSSR06	R: AATCTGCCTCTGGTAGTCCG	(GCC)7	58	In this study
CamSSR07	F: TCTCATCCCCATCTTTATCC	(TCC)7	58	In this study
CamSSR07	R: GTTCCCTGCTGCTGTTGTTA	(TCC)7	58	In this study
CamSSR08	F: TCACCAGTCACTTTCCCTCC	(AC)10	58	In this study
CamSSR08	R: CCACCAAAAGGCACAATACC	(AC)10	58	In this study
CamSSR09	F: CATCATCCATCAAACCGTCC	(AT)10	58	In this study
CamSSR09	R: GAAGGCACATTGGTTCTGGG	(AT)10	58	In this study

Primers	Na	Ne	PIC
478	7.00	4.12	0.72
SSR2	6.00	2.63	0.58
CamsinM11	4.00	1.85	0.43
CamSSR01	3.00	1.48	0.29
CamSSR02	4.00	2.97	0.62
CamSSR03	6.00	4.96	0.77
CamSSR04	6.00	2.99	0.62
CamSSR05	4.00	2.82	0.58
CamSSR06	4.00	1.76	0.41
CamSSR07	4.00	1.58	0.35
CamSSR08	6.00	4.96	0.77
CamSSR09	5.00	2.32	0.49
(mean)	4.92	2.87	0.55