Genetic diversity analyses ofLasiodiplodia theobromaeonMorus albaandAgave sisalanabased on RAPD and ISSR molecular markers

Abstract

Genetic diversity of 23 Lasiodiplodia theobromae isolates on Morus alba and 6 isolates on Agave sisalana in Guangxi province, China, was studied by using random amplified polymorphic DNA and inter-simple sequence repeat molecular markers. Results of two molecular markers showed that the average percentage of polymorphic loci of all isolates was more than 93%. Both dendrograms of two molecular markers showed obvious relationship between groups and the geographical locations where those strains were collected, among which, the 23 isolates on M. alba were divided into 4 populations and the 6 isolates on A. sisalana were separated as a independent population. The average genetic identity and genetic distance of 5 populations were 0.7215, 0.3284 and 0.7915, 0.2347, respectively, which indicated that the genetic identity was high and the genetic distance was short in the 5 populations. Average value of the gene diversity index (H) and the Shannon’s information index (I) of 29 isolates were significantly higher than 5 populations which showed that genetic diversity of those isolates was richer than the populations and the degree of genetic differentiation of the isolates was higher. The Gst and Nm of 29 isolates were 0.4411, 0.6335 and 0.4756, 0.5513, respectively, which showed that the genetic diversity was rich in those isolates.

Characterization of Lasiodiplodia isolates obtained from acid lime and Valencia orange in Egypt

“…The conidia of the pathogen were initially unicellular, hyaline ellipsoid to subovoid. Mature conidia were dark brown, bicelled, and thick walled and ellipsoid described by Honghui Xie et al 2016 Primer Name Sequence ISSR-1 5'-AGAGAGAGAGAGAGAGYC-3' ISSR-2 5'-AGAGAGAGAGAGAGAGYG-3' ISSR-3 5'-ACACACACACACACACYT-3' ISSR-4 5'-ACACACACACACACACYG-3' ISSR -5 5'-GTGTGTGTGTGTGTGTYG-3' ISSR -6 5'-CGCGATAGATAGATAGAT-3' ISSR-8 5'-AGACAGACAGACAGACGC-3' ISSR -9 5'-GATAGATAGATAGATAGC-3' ISSR -16 5'-GACAGACAGACAGACAAT-3'…”

“…To detection the polymorphism between the isolates, 9 ISSR primers (table 1) were used to amplify the DNA of 6 isolates (Table 1). ISSR PCR amplification accomplished according to Hong-hui Xie et al, (2016). The amplification was carried out in a 25-µl reaction volume containing 1-µl template DNA (approximately 30 ng/µl), 1 µl primer (10 μmol/l), 0.5 µl dNTP (10 mM /μL), 2.5 µl Taq Buffer, 2 µl 25 mM MgCl2, 0.2 µl Taq polymerase and 17.8 µl ddH2O.…”

“…The amplification products of ISSR PCR were size separated by electrophoresis in a 1.5% agarose gel containing ethidium bromide (0.5ug/ml) in1 × TAE buffer (40 mM Tris, 20 mM acetic acid and 1 mM EDTA) at 80 V for 120 min and 100 V for 180-200 min at room temperature, respectively Hong-hui Xie et al, 2016. The gels were visualized under UV light and photographed using a Gel Documentation System (BIO-RAD 2000). The sizes of amplified DNA fragments were estimated by comparison with 10 kb DNA ladder markers.…”