Aquaporins in the wild: natural genetic diversity and selective pressure in the PIP gene family in five Neotropical tree species

Table 2 Description of the PIPs fragments amplification conditions

	Species	Primer Name	Primer sequences (5' → 3')	T _a (°C)	Size (bp)	Genbank accession number
a.	All	PIP2H2.2	F: CTYGTYTACTGCACHGCY	64	850	-
		PIP2H6.1	R: CCVACCCARAADATCCAN
b.	Carapa guianensis	CguPIP1.1+0018	F: CGGCATTTCAGGTCATCTC	54	760	FJ709600
		CguPIP1.1-0780	R: CCAACCCAGAAAATCCAGTG
	Pachira quinata	PquPIP2.1+0017	F: GCCGGTATCTCTGGTGAGTG	64	650	FJ709598
		PquPIP2.1-0672	R: CCACGCCTTCTCTTTGTTGT
	Virola sebifera	VsePIP2.1+0032	F: CGCGTATCTCTCTCTTCAACG	59	750	FJ807641
		VsePIP2.1-0788	R: CACACGCACACACACAATG
	Eperua falcata	EfaPIP1.1+0043	F: CCCAGCAGTGACCTTCG	64 → 57 ⁽¹⁾	550	FJ807642
		EfaPIP1.1-0487	R: AACCAAGAACACAGCGAACC
		EfaPIP1.2+0040	F: CAACCCGGCTGTGACC	64 → 57 ⁽¹⁾	550	FJ807646
		EfaPIP1.2-0487	R: GCCAAATGGACCAAGAACAC
		EfaPIP2.1+0034 ⁽²⁾	F: GCACATAAATCCGGCAGTG	64 → 57 ⁽¹⁾	650	FJ807645
		EfaPIP2.1-0484 ⁽²⁾	R: CCGACCCAGAAGATCCAC

Legend: Primer names, primer sequences, annealing temperature, size of the sequenced fragment, accession number of the closest Arabidopsis thaliana BLAST match. Primer names are derived from clone name followed by a sign corresponding to the direction of the synthesis ("+" for "forward", "-" for "reverse") and a number corresponding to the position of the primer's 5' end relative to the clone's sequence. ⁽¹⁾ PCR touchdown: the first seven cycles with one degree decrease each cycle, from 64°C to 57°C. Other cycles at 57°C
⁽²⁾ For E. grandiflora, the same primers and conditions were used for PCR amplifications.

ISSN: 2730-7182