Abstract:
Total RNA was isolated from
Siraitia grosvenorii fruit by the method of modified Trizol, according to
S. grosvenorii fruit characteristics of rich phenols, polysaccharide, oil and proteins. The OD
260/280, OD
260/230, RNA integrity (RIN) and yield of the total RNA with this method were 2.01, 2.02, 9.50 and 260 礸·g
-1, respectively. The open reading frame (ORF) of dehydroascorbate reductase (DHAR), named as
SgDHAR, was cloned by rapid amplification of cDNA ends (RACE) and RT-PCR method from
S. grosvenorii. The GenBank accession number for this gene is KC907731. The
SgDHAR gene contains a full-length cDNA of 1 252 bp including ORF of 819 bp and encodes a predicted protein of 272 amino acids. The molecular mass is 30.217 7 kD and the isoelectric point is 8.76. Homology comparison showed that it shared 87% nucleotide sequence homology with
Cucumis sativus. Expression patterns using qRT-PCR analysis showed that
SgDHAR was mainly expressed in fruit and stem, followed by flower, and was lowest in root, while the expression level was 6.83 times in triploid than that in diploid. Therefore,
SgDHAR gene may be involved in abortion of triploid seedless
S. grosvenorii.