Novel Microsatellite Markers for Brazilian Mangrove Oysters ( Crassostrea gasar ) and their Cross-Amplification in Crassostrea rhizophorae

Baldez, Renata do Socorro Corrêa; Melo, Mauro André Damasceno; Sampaio, Iracilda; Tagliaro, Claudia Helena

doi:10.1590/1678-4324-2016160071

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Biological and Applied Sciences • Braz. arch. biol. technol. 59 • 2016 • https://doi.org/10.1590/1678-4324-2016160071 linkcopy

Novel Microsatellite Markers for Brazilian Mangrove Oysters ( Crassostrea gasar ) and their Cross-Amplification in Crassostrea rhizophorae

Authorship SCIMAGO INSTITUTIONS RANKINGS

A microsatellite CT/GT enriched genomic library was developed for Crassostrea gasar and twelve new polymorphic loci were isolated and characterized. The markers were successfully amplified from 25 individuals of Crassostrea gasar and 11 cross-amplified individuals of Crassostrea rhizophorae. There was no evidence of linkage between loci in either species.

Key words:
SSR; genetic markers; cupped oyster; population structure.

Authorship

person Renata do Socorro Corrêa Baldez

person Mauro André Damasceno Melo

school Universidade Federal do Pará, Campus Bragança, Laboratório de Conservação e Biologia Evolutiva, Bragança, Pará, Brasil. Universidade Federal do ParáBrazilBragança, Pará, Brazil Universidade Federal do Pará, Campus Bragança, Laboratório de Conservação e Biologia Evolutiva, Bragança, Pará, Brasil. school Instituto Federal do Pará, Campus Bragança, Laboratório de Biologia Molecular e Ambiental, Bragança, Pará, Brasil.Instituto Federal do ParáBrasilBragança, Pará, Brasil Instituto Federal do Pará, Campus Bragança, Laboratório de Biologia Molecular e Ambiental, Bragança, Pará, Brasil.

person Iracilda Sampaio

person Claudia Helena Tagliaro ^*

*Authors for correspondence:
emailtagliaro@ufpa.br

SCIMAGO INSTITUTIONS RANKINGS

Universidade Federal do Pará, Campus Bragança, Laboratório de Conservação e Biologia Evolutiva, Bragança, Pará, Brasil. Universidade Federal do ParáBrazilBragança, Pará, Brazil Universidade Federal do Pará, Campus Bragança, Laboratório de Conservação e Biologia Evolutiva, Bragança, Pará, Brasil.

Instituto Federal do Pará, Campus Bragança, Laboratório de Biologia Molecular e Ambiental, Bragança, Pará, Brasil.Instituto Federal do ParáBrasilBragança, Pará, Brasil Instituto Federal do Pará, Campus Bragança, Laboratório de Biologia Molecular e Ambiental, Bragança, Pará, Brasil.

Tables

Tables (2)

Thumbnail

Table 1 Locus name, repeat motif, primer sequences and size ranges (bp) of twelve microsatellite loci isolated from Crassostrea gasar and cross-species amplification with Crassostrea rhizophorae.

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Table 2 Characterization of twelve microsatellite loci isolated from Crassostrea gasar and cross-species amplification with Crassostrea rhizophorae, including locus name, number of alleles (n), annealing temperature (T), observed (Ho) and expected (He) heterozygosities, and probabilities of deviation from Hardy-Weinberg equilibrium (P _HW).

table_chartTable 1 Locus name, repeat motif, primer sequences and size ranges (bp) of twelve microsatellite loci isolated from Crassostrea gasar and cross-species amplification with Crassostrea rhizophorae.

Locus name	Repeat motif	Primer sequences (5'-3')	Size range (bp) C. gasar	Size range (bp) C. rhizophorae
CGA07	(CT)₂₃	GATACTCTCTCGCTCTCACAGAGTT TCGGGTGGGTAATTTAGAGGTA	164 - 180	166 - 176
CGG08	(AC)₁₀	CCGCACTAGTGATTCTCTTGCT CGTGCGGGTATTACTGTCTTGA	106 - 338	126 - 388
CGH05	(AC)₁₇(AT)₂₅ (AC)₅	AATTCACAAGTAGAATTCCCAAAGA AGTTGAAAAAGGAAAACAGGATGTA	170 - 296	166 - 214
CGB06	(TC)₇TG(TC)₁₄	GTAGAGGACATTCCCGAGAAGA CTCCACAGTCGCATACAAGGTT	206 - 286	212 - 268
CGH03	(CA)₈	GATCCAGCAGGTGTTAGGAGAT ACACTAAGCATCGGCTTGTTGG	160 - 226	162 - 206
CGA12	(CA)₉	CTTGATGCCGACCACTGTAATA TTACCTTGTTTGTGAGGAGAGGTC	180 - 188	180 - 184
CGB09	(CT)₁₆TT(CT)₇(GT)₁₇GG(CA)₈	CAGCGAGTCTTAGCAATCTCTC GGCTATGGGAGTCTCGATATAGG	358 - 410	338 - 406
CGE11	(GT)₄CG(GT)₂GC(GT)₉	GTAAGTGTAAAACGTCTTTGCTGTC TAGTGCAACCCGTGTTTATATGACT	180 - 202	184 - 202
CGD05	(TG)₁₁(AG)₂₁	TAGTCGATCGATCTCTCGTTCC TAGCGGTCTTTGACCAGTATCG	408 - 450	400 - 448
CGG06	(TC)₃TT(TC)₂₂(C)₁₀(CA)₇	CCGTTAAGGTGTCCATATACTGC GTAAGAAGGTGGTCGGGGAGTAA	214 - 264	218 - 256
CGH02	(GT)₁₄	AGCGTCTTTACGAGTTTGTTGC AGATGAAGACGTCACCGTTACAG	222 - 250	230 - 252
CGF03	(GT)₈	AAAGGTGAGATGATCGTAGGAG CGCGTGGACTAACTAAGACAACAG	400 - 452	400 - 436

table_chartTable 2 Characterization of twelve microsatellite loci isolated from Crassostrea gasar and cross-species amplification with Crassostrea rhizophorae, including locus name, number of alleles (n), annealing temperature (T), observed (Ho) and expected (He) heterozygosities, and probabilities of deviation from Hardy-Weinberg equilibrium (P _HW).

Crassostrea gasar (25 specimens)						Crassostrea rhizophorae (11 specimens)
Locus name	Number of alleles (n)	T (°C)	Ho	He	P _HW	Locus name	Number of alleles (n)	T (°C)	Ho	He	P _HW
CGA07	8	52.5	0.750	0.839	0.157	CGA07	6	52.5	0.818	0.793	0.190
CGG08	17	55	0.875	0.902	0.000*	CGG08	11	55	0.900	0.855	0.427
CGH05	19	60	0.870	0.922	0.129	CGH05	6	61	0.833	0.792	0.410
CGB06	22	64.5	0.833	0.941	0.027	CGB06	13	64.5	0.818	0.868	0.160
CGH03	16	62	0.720	0.882	0.000*	CGH03	10	61.5	0.444	0.877	0.000*
CGA12	5	51	0.667	0.718	0.629	CGA12	3	51	0.636	0.562	1.000
CGB09	16	60	0.917	0.923	0.224	CGB09	13	60.5	0.818	0.897	0.227
CGE11	10	60	0.680	0.790	0.167	CGE11	8	60	0.636	0.793	0.044
CGD05	15	60	1.000	0.900	0.463	CGD05	13	60	0.909	0.909	0.521
CGG06	21	60	0.840	0.927	0.024	CGG06	13	60	0.818	0.913	0.165
CGH02	13	60	0.875	0.884	0.418	CGH02	10	60	0.909	0.855	0.649
CGF03	6	64	0.522	0.622	0.078	CGF03	4	64	0.455	0.380	1.000

* Significant P _HW value after Bonferroni correction P < 0.005.

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