Identification Of Hybrids From Lilium By In Situ Hybridization

Cross breeding is an important means of lily breeding.Breeding lily varieties with various characters is an urgent task for lily breeding in China.The research group has been devoted to the hybrid breeding of lily.Through the conventional hybridization breeding methods and embryo rescue methods,more than 10000 hybrid hybrid progenies were obtained from more than 60 hybrid combinations.But these hybrid offspring may be false hybrids,so we need to identify the authenticity and decide their trade-offs.Due to the longer vegetative growth period,early hybrid identification can reduce the waste of manpower and material resources and improve the efficiency of breeding.Therefore,the identification of authenticity is very important for the breeding of new lily varieties.This study starts with molecular cytogenetics,and uses genomic in situ hybridization to identify early hybrids.The main results are as follows:(1)In this study,the hybrid parents of LA hybrid Lilium 'Menorca','Original Love' and 'Pollyanna' and 'Indian Diamond' were hybrid parent materials.Four combinations of cross breeding were carried out.‘Menorca'בPollyanna'(3x×2x)?‘Menorca'בIndian Diamond'((3x×4x)?‘ Original Love'בPollyanna'(3x×2x)?‘ Original Love'בIndian Diamond'((3x×4x).After embryo rescue,different number of progeny plants were obtained.(2)The probe preparation,the composition of hybrids and the key steps and details of the GISH technology applied to this group were explored,and the GISH platform of our group was initially set up.The method of making the chromosome making method is to take the method of pressing.The method of preparation of the probe is to bathe the extracted Lily DNA in the boiling water for 15 min.The preparation method of the sealing resistance is to extract the DNA boiling water bath 40 min of the lily.The hybrids were composed of 50% deionized formamide(Formamide),10%(W/V)glucan(Dextransulphate),2 x SSC(pH=7.0),0.25%(W/V)twelve alkyl sulfate(SDS),0.6-1.5ng.L-1 L-1 probe DNA and 25-100 ng L-1 sealing resistance system.The in situ hybridization time was about 2,2 and 0.1 * two.The method of step elution,first soaking and concussion,the detection of hybridization signals using two kinds of antibodies,20 g.Ml-1anti-dig-FITC and 20 g.Ml-1rabbit-anti-sheep-FITC,were detected.(3)GISH Hybrid Progenies of 'Menorca' and 'Pollyanna' hybrid offspring 16-8,'Menorca' and 'Indian Diamond' hybrid 16-11,'Original Love' and 'Pollyanna' hybrid descendants 16-13,'Original Love' and 'Indian' hybrid offspring 16-14 were identified for authenticity.The results showed that 4 hybrids are the parents of the son of generation.4 hybrids were all aneuploidy,and 3 hybrids had recombination chromosomes.Both 'Menorca' and 'Original Love' can produce fertile aneuploidy female gametes,can be hybridized as a parentand.And their female gamete meiosis is not strictly paired with 24 Asian chromosomes to form 2 valence bodies,and some large spores can be paired and exchanged to form recombinant chromosomes of the Asian lilies and musk lilies.The chromophore is used to achieve introgression between Asian Lily and musk Lily in the offspring.It's a good breeding material.