Inducement Of Seeds Germination And In Vitro Inoculation Of Cistanche Tubulosa (Schenk) R.Wight

In vitro morphology of seed germination and haustorium formation in Cistanche tubulosa(Schenk) R.Wight without the host was studied firstly,effects on seeds germination with various fore treatments(chilling, decortication, treatment with oxidant NaClO), extraneous plant growth regulator and conjunct culture of seeds with radicated host tube seedlings were investigated in order to search effectual methods of seed germination inducement .Since C.tubulosa's host Tamarix chinensis Lour is prone to rapidly propagate, effects of roots of C. tubulosa's tube seedling on seeds germination haustorium formation and parasitism were investigated, we elementarily determinate the nature of Germination stimulants in C.tubulosa. Then an system of in vitro rapid propagation of Tamarix chinensis Lour was established through tissue culture, seed coats have been peeled off before seeds were inoculated on root of tube seedling of Tamarix chinensis, thus an in vitro inoculation system of tube seedling of Tamarix chinensis Lour using seeds of Cistanche tubulosa (Schenk) R.Wight was developed .The results are as follows:1. Seeds began to geminate after two weeks. Radicle elongated out of the micropylar end of the soften endopleura, formed germ-tube-like organ, and then elongated gradually. Seventeen days later, elementary haustoria began to develop and several haustorial hairs were differentiated from thick ribbon epiderm which formed fossae among apex of elementary haustoria. Papillary protuberance known as papillae scattered on winkles of epiderm and in fossae. When they elongated till 4 mm , germ-tube-like organs ceased development. The process of decorticated seeds germination and haustorium formation is as follws:sprouting—protuberance(tapered shape) —formation of germ tube(slightness) —germ tube growing widely—intumescence in proximal end — attenuation in acropetal end.2.Various fore treatments(chilling, decortication, treatment with oxidant NaClO) and extraneous cytokinin play an active part in promoting seed germination rate,startup of germination in decorticated seeds with GA₃ treatment was ahead of schedule than control, treatment of nomal seeds with GA3 at high concentration can improve rate of germination remarkablelly. Stratified seeds were decorticated and soaked in 2%NaClO eight minutes,then they were sown on 1/2 MS medium solidified with agar supplemented with NAA 0.1mg/LN KT 0.5 mg/L and GA₃ 0.5 mg/L,seed germination rate may add up to 35.5%. Roots of tube seedling of Tamarix chinensis can increase seed germination rate remarkablely, seed germination rate may reach 31.6% when seeds were grown on substrate with radicated host tube seedlings.3. The system of in vitro rapid propagation of Tamarix chinensis Lour was established through tissue culture, then investigation was made on interaction between methylene blue and secretions of roots of host test tube seedlings in MS medium solidified with agar supplemented with 200 mg/L methylene blue. Methylene blue faded in culture medium with the presence of host roots, whereas control without the development of host roots did not fade. The experimental results indicated that a kind of deoxidized hydroquinone which functioned through a mechanism of the redox reaction was identified as germination stimulants of Cistanche tubulosa.4. Seed coats have been peeled off before seeds were inoculated on root of tube seedling of Tamarix chinensis, investigations on inoculation were carried through by stereo microscope and observations on infection were done by means of whole clearing method. Haustorium of Cistanche tubulosa and xylem of host Tamarix chinensis were jointed, results indicated that in vitro infection onhost roots with seeds of Cistanche tubulosa was achieved successfully. Thus cistanche tubulosa-Tamarix chinensis inoculation complex can be established through culture of seeds of Cistanche tubulosa together with radicated Tamarix chinensis Lour.