| To determine the suitable stages and hormones combinations for inducing callus of the processing tomato,anthers of two varieties processing tomato as material.Anthers were crushed and made tablets with acetic acid magenta,then observed spore development stage by stereomicroscope.MS medium supplemented with different concentrations of plant hormones such as 2,4-D,NAA,KT,6-BA,ZT,TDZ for anther culture to study the effects of different combinations on callus induction of processing tomato anthers.The test results are as follows:(1)Determination of anther microsporidium during processing of tomato :The color and size of tomato buds can be used as a simple and reliable indicator to determine the development period of microspore,and to process tomato CK8 and CK3.When the buds are small,completely closed,the sepals are tightly enveloped by petal,green,the buds’ longitudinal diameter is 2.44-2.96 mm,and the transverse diameter is 1.48-1.63 mm,most of the microspores are in the stage of pollen mother cells.When the flower buds are slightly larger and still completely closed,the sepals are still tightly packed with petals,showing a green color,the buds have a longitudinal diameter of 3.32-4.08 mm and a transverse diameter of 1.60-2.19 mm,most of the microspores are in the tetrad stage.When the petals were lower than the bracts,the bracts were green,the petal was light green,the buds’ longitudinal diameter was 3.99-5.26 mm,and the transverse diameter was 2.04-2.33 mm.Most of the microspores were in the mid-mononuclear stage.When the petals are slightly lower than the bracts,green,petal light green,yellowish,buds 5.34-6.05 mm in longitudinal diameter,and 2.25-3.07 mm in transverse diameter,most of the microspores are located on the edge of the mononuclear period.(2)Effects of different hormone combinations and concentrations on anther culture of processed tomato :Hormones play an important role in tomato anther culture.In MS medium,different concentrations of 2,4-D,NAA and different concentrations of KT,6-BA,ZT,and TDZ were added.The results showed that 0.5 mg/L 2,4-D+0.05 mg/L TDZ,0.1 mg/L 2,4-D+2.0 mg/L 6-BA,0.1 mg/L 2,4-D+0.05 mg/L TDZ is suitable for CK8,the callus induced combination of hormones and induction rate of anthers were all above 56 %,0.5 mg/L 2,4-D+0.1 mg/L TDZ was also a hormone combination suitable for callus induction of CK8 anthers,and the induction rate was 52 % the above.0.1 mg/L 2,4-D+0.1 mg/L TDZ was the most suitable hormone combination for CK3 callus induction with an induction rate of 58.04 %,0.1 mg/L 2,4-D+2.0 mg/L 6-BA,the induction rate for CK3 was the second highest,which was 55.05 %,the induction rate of 0.1 mg/L 2,4-D+1.0 mg/L 6-BA was also high for CK3,and the induction rate was up to 51 %.The inducing rate of CK8 by the combination of 2,4-D +TDZ was better than that of other combinations,while the induction of CK3 by the combination of 2,4-D + 6-BA was better than other combinations.The CK3 anthers were cultured on the induction medium MS +0.5 mg/L NAA +0.05 mg/L TDZ and subcultured MS +0.5 mg/L NAA +0.5 mg/L KT.Embryogenic callus was induced on differentiation medium MS + 0.2 mg/L NAA +0.5 mg/L KT.(3)Effects of genotypes on callus induction of processed tomato anthers :The callus induction rate of anther culture of different genotype materials was different.The callus induction rate of the two genotypes inoculated with the test were 18.63% and 14.19% in CK8 and CK3,respectively.(4)Effect of low temperature pretreatment at 4°C on callus induction of processed tomato anthers :Two CK8 and CK3 anthers of the two genotypes were pretreated at 4 °C for 2 d and 5 d before inoculation.The results showed that the two genotype-processed tomato anthers were pretreated at 4 °C for 2 d and 5 d at low temperature.The difference of induction rate was not obvious. |
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