Utilization Of Saccharum Related Genera Erianthus And Identification The Genuine Hybrids From The Cross Of Saccharum And Erianthus

With the financial support from "Hi-Tech Research and Development Program of China (863 program)" (2001AA241191) and "National Natural Science Foundation of China" (30170590), Analyses of ISSR and ITS markers, chromosome karyotype and mRNA different display were carried out to study on the genetic diversity among 25 clones of Erianthus sect. Ripidium, identification of genuine hybrids from the cross of Saccharum and Erianthus, and the different gene expression in salt torelance of Erianthus. Based on the cluster and principal component analysis on the ISSR and ITS PCR amplification from the tested clones of Erianthus, all the clones could be divided into four groups according to the nearest phylogenetic relationship. In most cases, clones from the same region were in the same group and only the clones in few regions could not be clustered into the same group. Nine F1 from the cross of Saccharum and Erianthus thirty-six BCl from the cross of hybrids of Erianthus (YC96-66,YC96-46) and CP84-1198 twelve BCl from the cross of YC95-41 and Neijian57-416 were identified using ITS-PCR with the primer pairs of EF1/ER1 and EF2/ER2. The results from ITS-PCR identification for F1 hybrids of Erianthus is accorded with isozyme identification that was reviewed by SHEN Wan-kuan. However, most of fifteen clones which had been considered as the genuine hybrids from the cross of Saccharum and Erianthus, were not genuine hybrids in this research.The cytogenetic studies on the hybrid and backcross progenies of the five tested samples of sugarcane and Erianthus showed that the somatic chromosome number of progeny Ya 96-66 originated from Badila (female parent, 2n=80) with Erianthus 95-105 (male parent, 2n=60) was 2n=70, following the crossing law of n+n; the crossing between Ya 96-66 (female parent) and CP84-1198 (male parent, 2n=120) also followed the same genetic law and the somatic chromosome number of their progeny, Ya 01-3, was 2n=105. These results demonstrated that sugarcane and Erianthus was crossed in the way of n+n.Two anchored primers with "G" base and ten random primers for total 20DD-PCR reactions were carried out on mRNA different display in salt torelance of Erianthus. Nine differential cDNA bands were amplified. However, when they were reamplified, only four bands were tested to be positive. The diffenential cDNA fragments were purified and inserted into PMD-T vector. The cloned cDNA fragment were sequenced and blasted in GeneBank. No highly homologous sequences in GeneBank indicated the differential clone may be new genes from Erianthus subjected to salt stress. They will be confirmed with Northern and Southern blotting.