Studies On Crossing-compatibility And Embryo Culture Of Hybrids Of Pistacia Vera L.

The flower bud differentiation，male and female gametophyte development male andthe optimal conditions of pollen germination for Pistachio (Pistacia vera L.)and PistaciaChinensis Bunge were studied by using of anatomy, palynology and morphological，physiological and biochemical analysis. In order to understand the the key period of hybridembryo abortion characteristic and establishing hybrid embryo rescue technology system,the process of “Pistacia vera and Pistacia chinensis” cross, the hybridization compatibility，the fertilization and the embryo abortion were observed by using fluorescence microscopyand in vitro embryo technology. The adaption of F1hybrid such as salt tolerance and coldresistance ablity were also evaluated in the paper. The results are as follows:1. The discipline of pistachio male plants of T-491and Pistacia Chinensis Bungeinflorescence growth earlier，since March20，inflorescence growing season lasts about15d，and the female plants Kerman late inflorescence growth begins on about March28，while the later on about20d. Early flowering female plants Kerman begins about on April9to10，and the phenophase of Kerman and male plant T-491overlap in the April10to12，only3d. The phenophase of Pistacia Chinensis Bunge and Kerman flowering overlap inthe April10to14，about5d.2. The microsporogenesis，megasporogenesis，and development of the male and femalgametophytes of Pistachio were observed by paraffin section technique to test whetherthere is any obstacle of reproduction in the process of sexual reproduction.The resultsshowed:(1) The wall of anther consists of epidermis endothecium，middle layers andtapetum from out to inside，tapetum cell is of glandular tapetum.(2) Cytokinesis duringmeiosis of the microspore mother cell was simultaneous type，and tetrads were isobilateral.(3) Mature pollen grain was2-cell type with one or three germ pores (4) There is an ovarythat contains one ovule，ovules occur at the base of ovary. The ovule was anatropous，bitegmic and crassinucellate. The development of embryo sacs belonged to polygonumtype.（5）Some of female gametophyte was abnormal.3. The pollen grains were long-globous existing except with T’11pollen. The exineornamentation were all reticulate sculpture. The shape of T-11’s mesh was foveolae whilethe others’ were round or approximately round. The exterior of the T-491’pollen has a fewwhite grains and that of the other five breeds had a lot of white grains. The shape of T-11’sgerminal aperture was furrowed，that of Peter was round and that of the T-491，the PistaciaChinensis Bunge，UCB-1and Gan Gu were oval. The order of pollen size is: Peter＞thePistacia Chinensis Bunge＞T-491＞T-11＞Gan Gu＞UCB-1.4.The bourgeon pollen of Pistachio and Pistacia Chinensis Bunge suitable medium was composed of15%sucrose，100mg/L boric acid，cultivated at25℃；The suitableconcentrations of GA3，2,4-D for the pollen germination and tube growth of Pistachio andPistacia Chinensis Bunge were25mg/L and5mg/L which was the pollen germination rateand the length of pollen tube reached to the largest. The concentration of NAA was0.5mg/L，pollen germination rate reached to the largest.When the temperature or theconcentration were rised or reduced，the pollen germination rate and the length of pollentube would be restrained in a certain degree. Pollen activity of SOD，POD and CATactivity after storage present first increased and then decreased.5. Using fluorescence microscopy observed that the the hybrid combinations stigmaattached after pollination and germination of pollen count is less than the contrast，pollengermination has lagged far behind，and the number of pollen tubes reach the style andovary less than that of the contrast. Intergeneric Cross of stigma and style in a largenumber of irregular callose deposition，pollen tube bending，swelling thicker phenomenon，24h after pollination，a handful of pollen tube enters the ovary at24h，and finishedfertilization at72h after hybridization. Abnormal pollen tube growth and callose depositingafter pollination may be caused by the incompatibility of intergeneric cross betweenPistachio (Pistacia vera L.)and Pistacia Chinensis Bunge.6. Embryo abortion stage and rescue system of hybrids were studied in the distanthybridization between Pistachio(Pistacia vera L.) and Pistacia Chinensis Bunge. Theresults showed: Intergeneric cross has embryo abortion phenomenon after pollination.notcompletely degraded integument and funicle browning spots，necrotic part of the extensionto the embryo sac in fruit. Pistachio hybrid embryo rescue embryo appropriate age100to120days after pollination，the development of hybrid embryo began from80days afterpollination，and the critical period for the hybrid embryo abortion was from the80to100days.The suitable multiplication medium for immature embryo seedlings was DKW+2.0mg/L6-BA+0.05mg/L IBA+0.3g/L LH+1.5g/L PVP，with which the multiplicationcoefficient of3.47and the seedling height of3.32cm were obtained. The suitable rootingmedium was1/2WPM+2.0mg/L IBA+0.05mg/LNAA with a rooting rate of80%.7. The adaptive initial analysis showed that the hybrid F1generation of resistancedisease and cold hardiness better than the two parents. The curve of diurnal variation of Pnin Hybrid F1showed two peaks.with obvious midday depression of photosynthesis，netphotosynthetic rate,stomatal conductance，intercellular CO2concentration and transpirationrate and Fo，Fv/Fm and Fv/Fo were higher than that of two parents. Artificial lowtemperature process，the three tree twigs water loss rate has the trend gradually increased；SOD，POD activity first increased and then decreased trend and MDA content showedincreased；with the treatment temperature the reducting，the soluble sugar content hasincreased trend. 8. Research on salt tolerance of the Pistacia vera L.and Pistacia chinensis BungeSeedlings show that（:1）With the increase of NaCl stress concentration，the cell membranepermeability，SOD activity gradually increased；MDA content has first decreased thenincrease in Pistacia vera L.and Pistacia Chinensis Bunge leaves，while POD activitydecreased;APX activity in Pistacia Chinensis Bunge leaves and GR activity in Pistaciavera L.has first increased and then decreased trend.（2）the content of Chl·a and Chl·b inPistacia vera L.and Pistacia Chinensis Bunge leaves were decreased while the Chla/b ratiowas enhanced of Pistacia vera L. by NaCl stress. The Chla/b ratio of Pistacia ChinensisBunge leaves has decreased，still significantly higher than the control; net photosyntheticrate(Pn)，stomatal conductance(Gs) of leaves significantly decreased.The decreased of Pnwas mainly attributed to stomatal limitation when the concentration of NaCl was lowerthan0.3%and to non-stomatal limitation when the concentration was higher than0.3%（3）With the increase of NaCl stress concentration，Trapped energy flux per RC（TRO/CSO），Electron transport flux per RC（ETO/CSo），Density of RCs（RC/CSO），Yields or fluxratios(φPo，φo�'�φEo) decreased，and Absorption flux per CS(ABS/CSO)， quantum yieldfor energy dissipation (φDo) increased.(4) With the increase of NaCl stress concentration,the content of Na+，Na+/K+ratio were increased，the content of K+were increased firstlyand then decreased in Pistacia vera L.and Pistacia Chinensis Bunge leaves.And with theincrease of NaCl concentration，the content of Ca，Mg，Fe，Zn content graduallydecreased.(5) When the concentration was higher than0.3%，the extracellular resistivity,relaxation time and distribution coefficient of relaxation time decreased consistently.