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Studies On In Vitro Organogenesis And Its Anatomy And The Changes Of The Endohormones Contents In Pinus Elliottii

Posted on:2010-05-21Degree:MasterType:Thesis
Country:ChinaCandidate:T Q XuFull Text:PDF
GTID:2143360275985137Subject:Garden Plants and Ornamental Horticulture
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Pinus elliottii is the main greening and forestation tree species in the south of China. Tissue culture is now the effective way for intermediate propagation. In this experiment, mature zygotic embryos were used as experimental materials to carry out the adventitious bud proliferation and plant regeneration. Also the further callus culture and the related anatomic obersavation was carried out. Besides, changes of endogenous hormone contents during bud proliferation were studied. The main results were described as follows:1 Studies on the adventitious bud proliferation of Pinus elliottiiAdventitious buds of Pinus elliottii were induced with the apical buds as explants. The influences of plant growth regulators, basal medium, hormone proportion, organics concentration etc. on proliferation of adventitious buds were experimented. The results showed that DP+6-BA 4.00 mg·L-1+NAA 0.05 mg·L-1 was the best medium for proliferation, the supplement of 10 g·L-1 inositol or 300 mL·L-1 coconut milk could promoted the proliferative effect of adventitious buds.2 Studies on callus culture, adventitious bud differentiation and anatomical observation of Pinus elliottiiCallus induction, growth and differentiation were studied with the mature zygotic embryos, sprouts, hypocotyls and radicle of slash pine as explants. The results indicated that the mature zygotic embryo was the best explant. TX + NAA 1.0 mg·L-1 + 6-BA 0.3 mg·L-1 and TX + 2,4-D 2.0mg·L-1 + NAA 0.5 mg·L-1 + 6-BA 2.0 mg·L-1 were the best media for callus cultivation and differentiation respectively, in which the induction and differentiation rates were both higher. Browning could not be inhibited in the experiment. It could only be reduced in the medium supplemented with Vc 0.05g·L-1 or PVP 1.00g·L-1 and by shortening subculture cycle. Different culture stage needed different intensity of illumination. The adventitious buds differentiated from callus had a low survival rate and were difficult to elongate. The histiocyte texture and difference were studied with callus as materials. Different kinds of callus had different texture of histiocyte. Only the white and compact callus had the capacity of differentiation. Meristemoid was the identification of organic callus. And the meristemoid formed near the surface of callus, so it belonged to the external origin.3 Studies on rooting of single shoot of Pinus elliottiiThe medium composed of both solid and liquid was used for rooting test. Results showed that the higher rooting rate was abtained in the solid DCR medium without major elements that was supplemented 20 days after culture.[0] Taking perlite and vermiculite as culture substrates instead of agar could not promote the rate of rooting. The supplement of different concentrations of ABT (0.25-4.0 mg·L-1) in the rooting medium had no obvious effect.4 Studies on the changes of endohormones contents during adventitious bud proliferation of Pinus elliottiiWith the adventitious buds induced from apical buds as explants, two different methods for adventitious bud proliferation and the changes of endogenous hormones contents in different proliferation stages were studied (ELISA). The results showed that with the proliferation generation increased, the proliferation coefficient declined in the way of adding 6-BA continuously and changed in a wave-like manner in the way of adding 6-BA in the intervening generation. In both of the two proliferation ways, the levels of endogenous ABA, IAA were both higher, and the change range of IAA was greatly. However, the levels of endogenous GA3 and ZRs were lower and their content changes were not obviously. Endogenous hormones contents of the two proliferation ways showed some differences at different proliferation stages. The levels of higher IAA and lower ZRs cut down the proliferation capacity. In summary, adding 6-BA in the intervening generation was the best way for adventitious bud proliferation of Pinus elliottii.
Keywords/Search Tags:Pinus elliottii, proliferation, adventitious root, callus, observation on anatomy, endogenous hormones, ELISA
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