| It’s the most efficient measure to get a higher sugarcane production level by breeding drought-resistant, disease-resistant and strong ratooning varieties. Germplasm innovation is the key for breeding breakthrough variety. Development and utilization of Erianthus arundinaceum, a closely related genus of Saccharum is the the most important thing for the Germplasm innovation of sugarcane. The hybriding utilization of Erianthus arundinaceum make slow progress because of the infertility and unclearness of the genrtic of the chromosome. Chromosome karyotype analysis was carried out with the F1and BC1of the progenies of sugancane and Erianthus arundinaceus and their parents, GISH analysis was carried out with the F1and BC1, inorder to discover the genetic of Chromosome. Microarray of sugarcane was used to hybridize with the cDNA from the different fertility clones or parents of BC1crossed from E.arundinaceus and sugarcane. The fertility-related genes were screened from the large number genes based on the bioinformatics analysis and microarray. The results were showed as follows:(1) Chromosome karyotype analysis show that the somatic chromosome number of E. arundinaceus Hainan92-105and Hainan92-77was both2n=60=60sm, belonging to type1A. The other five tested clones belonged to type2B. The somatic chromosome number of S. officinarum Badila was2n=80=70m+10sm, commercial variety CP84-1198was2n=120=114m+6sm(4SAT), The F,s YC96-66and YC96-40that originated from Badila with E. Arundinaceus were both2n=70=68m+2sm; Clones derived from the same cross of YC96-40(female) and CP84-1198(male) had different chromosome number, YC01-21had2n=104=100m+4sm chromosomes, YC01-134had2n=114=114m, YC01-36had2n=132=130m+2sm chromosomes.(2) The reslut of GISH show that The chromosomes of F,s YC96-66and YC96-40that originated from Badila with E. Arundinaceus were both consist of30chromosomes from E. Arundinaceus and40from S. officinarum. The chromosomes of BC1YC01-134was consist of29chromosomes from E. Arundinaceus and85chromosomes from Saccharum.spp. YC01-36was consist of36chromosomes from E. Arundinaceus and96chromosomes from Saccharum.spp. It’s the first time to find that chromosome transmit by the way of more than the somatic chromosome number. The phenomenon of distribut of chromosome and chromatin was both find in the four progenies of Saccharum.spp and E. Arundinaceus. The chromosomes of E. Arundinaceus always gather togather, but not so serious. There are one and two translocation chromosome in YC01-134and YC01-36respectively.(3)The results of karyotype analysis and GISH indicated that the hybrids originated from Badila (S. officinarum) and E. arundinaceus followed the chromosome transmission pattern of n+n. and the chromosome transmission of BC1were more complex, YC01-134with the genetic pattern of2n+n and YC01-36with another special kind of genetic pattern. The phenomenon of "Disequilibrium hybridization" was found in BC1.(4)4kinds compare of microarray expression profile between2fertility clones and2sterility clones show that14genes expressed4times and32genes expressed3times as Up-regulated genes;12gene expressed4times and34genes expressed3times as Down-regulated genes. There are92genes expressed Up-regulated or Down-regulated more than3tims. Three differentially expressed ESTs with ratio more than2were selected randomly for analysis of quantitative real-time RT-PCR.The similar results from microarray and Q-PCR indicated the reliability and authenticity in the microarray analysis.(5) The92Up-regulated and down-regulated ESTs gene sequences were identified and blasted at NCBI according to gene ID. A number of fertility genes including SR (serine-arginine repressor protein), JLP (jasmonate-induced protein), Prm3and the Retrotransposon were screened from92genes. These genes control the flower organ building and the property of flower meristem, play their part in the fertility alteration。They play an important role in the pollen development, anther dehisence and development stages control.(6) Discussion was done to explain the chromosome composition of YC01-36, the reason may be attribute to that YC96-40is a hybrid with many E. arundinaceus chromosomes in S. officinarum cell, this cause the disturbance of metabolism, male sterile, poor fertility as femail also. Only very few female gamete with perfact chromosome combination or have more than2n chromosomes number caused by abnormal meiosis can put through the process of fertilization, embryonic development, germinate and growh, another reason may be causes by the the out-sync in the zygot with2n+n, so the chromosomes double partly. And these progeniese were easy to be find for few population in one cross. |
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