Studies on the cytological and histological of somatic embryogenesis of Hevea brasiliensis
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TextPublication details: IRRDB Conference: NR Industry: Responding to Globalization 7-8 September 2004, Kunming International Convention & Exhibition Center, ChinaSubject(s): Summary: The somatic embryoids were induced in vitro with the explants, anthers of Hevea brasiliensis, Haiken 2, cultured on modified MB medium and MS basic medium with growth hormones (Chen Chenghua et al., 1978). During callus culture, calli were sampled every week from the phase of the anthers inoculation to torpedo embryoid, and sampled aperiodically according to the development status of somatic embryoid to observe systematically that the histology of the originating mode and development process of the somatic embryos of Hevea by microscope, and it were analyzed that each phase of somatic embryos generating and the cause of abnormal embryos formation of Hevea. And the following results were observed. 1. Somatic embryos of Hevea anther generated mainly in indirect mode. Most of the somatic embryo of Hevea originated from callus tissue, the several layers cells near by the external callus and surrounded by tannin cells that was the single embryonic original cell origination. 2. The process of somatic embryo development. A single embryonic original cell, which had polar, first divided unequally into two cells, one anear the external callus was the apical cell, and the other closed with the internal callus was the basal cell. And then, the apical cell successively divided vertically respectively to form 2-celled, 4-celled, and multicellular proembryos and then developed into globular embryoid, heart-shaped embryoid, torpedo embryoid, cotyledonary embryoid. And the basal cell divided transversely to form suspensor structure, which consisted of single row cells at the beginning and many rows cells in globular embryoid period. The suspensor structure existed even in the heart-shaped and cotyledonary embryoid period. During the embryonic original cell devided into multicellular proembryos, the cell wall of the external cell was thickened to form a clear cell boundary to separate itself from the ambient cells, namely the phenomenon of isolation. The isolation could strengthen its polarity. Polarity established and isolation were the keys to the somatic embryo normal differentiation in Hevea tissue culture. 3. The embryoid phase from heart-shaped to torpedo-shaped was the key period during the somatic embryogenesis in Hevea. It was easily to induce to globular embryoid or heart-shaped embryoid, but only a few of them further developed into normal embryoids and finally formed normal plantlets and most of embryoids stayed at the globular embryoid or heart-shaped embryoid period and could not develop into normal embryoid, such as torpedo embryoid, but abnormal embryoids. 4. There were different abnormal embryoids in each developmental phase. The abnormal embryoids in the period of globular embryoid included the following three kinds: (i) no suspensor; (ii) no protoderm, (iii) the abortive globular embryoid. During normal or some abnormal embryoids globular embryoids developed into heart-shape embryoid and torpedo embryoid, some abnormal embryoids occurred and could be described as following three kinds: 1) no shoot apical meristem; 2) procambium developed abnormally; 3)abnormal shape, such as joint-embryoids, or finger-like, or leaf-like. Some abnormal embryos could die slowly in the heart-shape embryoid phase and torpedo embryoid phase, but some could still develop into abnormal plantlets in the cotyledonary embryoid phase. These abnormal plantlets were mainly assorted into two kinds: one was that the root-part developed normally but the stem-part abnormally. The stem-part was gobbet-like, or joint-embryoid, or cup-like, or trump-like, or leaf-like. The other was that the root-part and the stem-part were both abnormal.
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The somatic embryoids were induced in vitro with the explants, anthers of Hevea brasiliensis, Haiken 2, cultured on modified MB medium and MS basic medium with growth hormones (Chen Chenghua et al., 1978). During callus culture, calli were sampled every week from the phase of the anthers inoculation to torpedo embryoid, and sampled aperiodically according to the development status of somatic embryoid to observe systematically that the histology of the originating mode and development process of the somatic embryos of Hevea by microscope, and it were analyzed that each phase of somatic embryos generating and the cause of abnormal embryos formation of Hevea. And the following results were observed. 1. Somatic embryos of Hevea anther generated mainly in indirect mode. Most of the somatic embryo of Hevea originated from callus tissue, the several layers cells near by the external callus and surrounded by tannin cells that was the single embryonic original cell origination. 2. The process of somatic embryo development. A single embryonic original cell, which had polar, first divided unequally into two cells, one anear the external callus was the apical cell, and the other closed with the internal callus was the basal cell. And then, the apical cell successively divided vertically respectively to form 2-celled, 4-celled, and multicellular proembryos and then developed into globular embryoid, heart-shaped embryoid, torpedo embryoid, cotyledonary embryoid. And the basal cell divided transversely to form suspensor structure, which consisted of single row cells at the beginning and many rows cells in globular embryoid period. The suspensor structure existed even in the heart-shaped and cotyledonary embryoid period. During the embryonic original cell devided into multicellular proembryos, the cell wall of the external cell was thickened to form a clear cell boundary to separate itself from the ambient cells, namely the phenomenon of isolation. The isolation could strengthen its polarity. Polarity established and isolation were the keys to the somatic embryo normal differentiation in Hevea tissue culture. 3. The embryoid phase from heart-shaped to torpedo-shaped was the key period during the somatic embryogenesis in Hevea. It was easily to induce to globular embryoid or heart-shaped embryoid, but only a few of them further developed into normal embryoids and finally formed normal plantlets and most of embryoids stayed at the globular embryoid or heart-shaped embryoid period and could not develop into normal embryoid, such as torpedo embryoid, but abnormal embryoids. 4. There were different abnormal embryoids in each developmental phase. The abnormal embryoids in the period of globular embryoid included the following three kinds: (i) no suspensor; (ii) no protoderm, (iii) the abortive globular embryoid. During normal or some abnormal embryoids globular embryoids developed into heart-shape embryoid and torpedo embryoid, some abnormal embryoids occurred and could be described as following three kinds: 1) no shoot apical meristem; 2) procambium developed abnormally; 3)abnormal shape, such as joint-embryoids, or finger-like, or leaf-like. Some abnormal embryos could die slowly in the heart-shape embryoid phase and torpedo embryoid phase, but some could still develop into abnormal plantlets in the cotyledonary embryoid phase. These abnormal plantlets were mainly assorted into two kinds: one was that the root-part developed normally but the stem-part abnormally. The stem-part was gobbet-like, or joint-embryoid, or cup-like, or trump-like, or leaf-like. The other was that the root-part and the stem-part were both abnormal.
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