Assessment Of Genetic Of The Main Introduction Acacia Spp.in Fujian And Preliminary Study On Hybrid Testing

The paper uses the RAPD heredity analysis to 20 kind of introduction acacia spp. in Fujian and uses the ISSR heredity analysis to 19 kind of Acacia spp.,and then observes submicrostructure of 10 kind of Acacia spp. pollen through the scanning electron microscope, in the above conclusion, carries on the Acacia spp.inter species crossbreeding experiment as well as carries on the blood relationship using the ISSR technology to a hybrid combinationThe results are as follows:1．The reliable RAPD analysis system is established: the reaction volume is 20ul.Reaction mixture consists with13．72 ulddHo,2 .0mmol/LBuffer(with Mg),1．2mmol/L of each dNTP,0．28 mmol/L Taq polymerase,1．2 mmol/L random Primer,1．6 ul DNA.RAPD program is: 2 min at 94℃for predenaturation, then 38 cycles of 30sec at 94℃for denaturation, 1min at 38℃for annealing, 90sec at 72℃for extension, finally extension at 72℃for 7min.2．Fourteen primers used in twenty Acacia spp. Selected from 100 primers test on the base of number and frequency of polymorphisms .With the 14 primers, 193 bands are polyporphic (96．89%) .3．Carring on the cluster to the RAPD data is possible to know that, acacia fimbriata and acacia podalyriifolia gather first in the same place, next is acacia irrorata and acacia glaucocarpa, their different coefficient respectively is 0．15,0．22, the corresponding heredity also is 0．15,0．22．Acacia mangium hybrid 4 finally gets together, occuping the cluster chart periphery; When the value levels on D=0．43, its junction divides 20 kinds of acacia spp.into one big class group, one minor sort, two independent kinds.Namely, the first big kind is acacia aulacocarpa,acacia fimbriata,acacia podalyriifolia,acacia melanoxylon,acacia implexa,acacia auriculiformis,acacia leptocarpa,acacia confusa Merr,acacia crassicarpa,acacia mangium,acacia mangium hybrid 1,acacia clunii and acacia floribunda; The second kind cut includes acacia irrorata,acacia glaucocarpa,acacia decurrens,acacia neriifolia and acacia farnesiana; The third kind has acacia cincinnata ; The fourth kind only has acacia mangium hybrid 4．4．The reliable ISSR analysis system is established: the reaction volume is 10ul.Reaction mixture consists with 7．15 ulddH0,0．75mmol/LBuffer(with Mg),0．1mmol/L of each dNTP,0．1 mmol/L Taq polymerase,0．9 mmol/LPrimer,1．0ul DNA.ISSR program is: 2 min at 94℃for predenaturation, then 35cycles of 60sec at 94℃for denaturation, annealing based on primers for 1 min, 120sec at 72℃for extension, finally extension at 72℃for 7min.5．Nine primers used in nineteen Acacia spp. Selected from 50 ISSR primers test on the base of number and frequency of polymorphisms .With the 9 primers,93 bands are polyporphic (97．8 % )6．Seeing from the ISSR cluster chart, acacia irrorata and acacia glaucocarpa gather first in the same place, the different coefficient is 0．20, the corresponding heredity also is 0．20, then they gather with acacia decurrens in same place; Next , acacia mangium and acacia mangium hybrid 1 gather in the same place, the different coefficient is 0．27, the corresponding heredity also is 0．27; acacia cincinnata finally gets together, occuping the cluster chart periphery.When the value levels on D=0．50, its junction divides 19 kinds of acacia spp.into two big class groups, two independent kinds. Namely,the first big kind is acacia fimbriata,acacia podalyriifolia,acacia auriculiformis,acacia irrorata,acacia glaucocarpa,acacia decurrens,acacia neriifolia and acacia farnesiana; the second kind cut includes acacia floribunda,acacia melanoxylon,acacia implexa,acacia leptocarpa,acacia confusa Merr,acacia crassicarpa,acacia mangium,acacia mangium hybrid 1 and acacia clunii; the third kind has acacia cincinnata; the fourth kind only has acacia aulacocarpa. The classified result of ISSR analysis and the classified result of RAPD analysis are close, but incompletely same.7．Using scanning electron microscope to observate and comparing 10 Acacia spp. pollen shapes, indicate that 10 Acacia spp. pollen shapes are highly consistent, 16 syntheses, the pollen arrangement stands by the central 8 grains of pollen, each each is 4 grains, the boundary for the single layer 8 grains of pollen, the boundary is clear, 16 syntheses are oblate, the side view is ellipse.The single grain of pollen is rectangle or near rectangle, Acacia spp. pollen is possibly non ditch class.8．Through the set of bag pollination to Acacia spp. for the crossbreeding experiment, obtains 7 hybrid combinations to have pod, in which the acacia mangium hybrid 4×acacia crassicarpa combination, obtains a hybrid seedling to be planted in the greenhouse; In the acacia aulacocarpa×acacia auriculiformis combination, obtains the pod also in the maturity, other hybrid combinations also obtain pods and somemature seeds. They indicate that the inter species hybridization of acacia spp.isfeasible.9．Using the ISSR technology to the parents and filial generation pod which come fromacacia aulacocarp×acacia auriculiformis for the molecular appraisal.The result mayappraise that this hybrid pod is initially hybrid.