Study On The Hybrid And Parents DNA Fingerprint From F₁ Seeds

The genuineness and purity test of seeds is important on maize (Zea maysL.)production. It was necessary to ask claimant to provide parents seeds to be relationduring the maize purity and genuineness identification in the past. But, now, for toprotect variety patent and trade private, the claimant will not or can not provideparents seeds in the purity and genuineness identification, which make difficult fortester. At the same time, during the nurturing hybrids and producting seeds, aphenomenon can not been dencied that some inbreds used by others under withoutthe permission of the inbreds'breeders or owners. It is not usually easy to protect theinbreds' breeders or owners' right under this condition. It is very that the pericarptissue of F1 seeds have the same genetic background with female inbred tissue. Thispaper established a technique of obtaining the parents and hybrid DNA fingerprintunder without provided parents seeds based on SSR markers by F1 seeds.The resultswere summarized as follows: 1. The method of maize pericarp isolation was decided. After compared andanalysized ,the NaOH soaking chosed to use in the research.The method ofexacting DNA from the pericarp and kernel tissue and PCR system based SSRmarker were optimized . 2. Thirty out of 162 pairs of SSR primers were selected, which gaveclear ,repeated, stable profiles in the 60 inbred lines.30 primers produced 119polymorphic alleles. The average number of allele per SSR loci was 3.9 with a rangefrom 2 to 7. The size of fragments ranged from 73 to 527 bp. The polymorphicinformation content (PIC)for the SSR loci varied from 0.24 to 0.82 with an average of 2新疆农业大学硕士学位论文0.56.Five single hybrids ,Jingke9,Jingke2, Nongda108, Jingzao13, Tangkang5 andtheir parents profiles by thirty pairs of SSR primers. 3. The DNA of every sort' male ,female,hyrid pericarp,hybrid profiled by theparents complementary primers, respectively. Hybrids' pericarp fingerprintsobtained,Jaccard analysized showed:the similarity coefficient is 1 between pericarpfingerprint and female fingerprint and further to prove both have same geneticbackground.The 5 male seeds and 4 female seeds was accurly identificated from 94mixed parents seeds with hybrid pericarp fingerprint. According to the informationof pericarp fingerprint, the average result of purity test of 10 Nongda108 hybrids is92.5%,which is nearly equal to the result of plant identification 92%.It can concludedthat the pericarp fingerprint to be puriety test relation is reliable. 4. The accuracy and reliable pericarp fingerprint as relation in the maize puritytest and inbreds genuineness indentification's practical syetems were established.