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Echevarría-Machado I Sánchez-Cach LA Hernández-Zepeda C Rivera-Madrid R Moreno-Valenzuela OA 《Molecular biotechnology》2005,31(2):129-135
A protocol is described for rapid DNA isolation from Malvaceae plant species and different tissues of Bixaceae that contain
large amounts of polysaccharides, polyphenols, and pigments that interfere with DNA extractions. The method is a modification
of Dellaporta et al. The current protocol is simple, and no phenolchloroform extraction, ethanol, or isopropranol precipitation
is required. The method is based in the incubation of soluble DNA with silica, mix in batch during the extraction. The procedure
can be completed in 2 h and many samples can be processed at the same time. DNA of excellent quality was recovered and used
for polymerase chain reaction (PCR) amplification, restriction enzyme digestion, and Southern blot analysis. The method was
used with healthy Bixa orellana and virus-infected Malvaceae plants. 相似文献
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The aim of the present work was to evaluate the feasibility of mannose as a selection system for the future genetic transformation of annatto (Bixa orellana L.). Hypocotyl segments, inverted hypocotyls and immature zygotic embryos were inoculated onto a Murashige and Skoog's medium supplemented with B5 vitamins, 87.6 mM sucrose and mannose in different combinations, 2.8 g dm?3 Phytagel®, and 4.56 μM zeatin (organogenesis) or 2.26 μM 2,4-dichlorophenoxyacetic acid and 4.52 μM kinetin (somatic embryogenesis). Annatto explants did not regenerate on medium with mannose as the only carbon source when inverted hypocotyls and immature zygotic embryos were used. However, organogenesis (5 % frequency) occurred exclusively in hypocotyl-derived explants nearest to the crown (collar) of the seedlings. No further shoot development was achieved. Therefore the substitution of sucrose by mannose inhibited both organogenesis and embryogenesis, and thus the employment of mannose could constitute an alternative selective agent in protocols for genetic transformation of this species. 相似文献
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Marie-Claire D'Souza Madhuri Sharon 《In vitro cellular & developmental biology. Plant》2001,37(2):168-172
Summary A protocol for in vitro propagation of Bixa orellana is described. Plants were regenerated from shoot apex and nodal explants on B5 medium supplemented with 4.9 μM 2-isopentenyl adenine. The multiplication factor of shoot apex explants was higher (nine shoots per explant) than that of
the nodal explants (five shoots per explant). Regardless of the position of the nodes, all the nodal explants gave similar
responses. However, the size of the nodal explant was an important factor in producing multiple shoots: 0.5 cm nodal explants
produced the maximum multiple shoots. Regenerated shoots from shoot apex explants rooted best on MS medium supplemented with
0.05 μM α-naphthalene acetic acid (NAA). whereas shoots regenerated from nodal explants needed 2.7 μM NAA for rooting. Eighty per cent survival of in vivo transferred plants occurred on the best potting substrate, coco peat. Since the multiplication factor was nine per explant,
this protocol can be use for commercial microprogation. However, the regeneration capacity declined after 10 subcultures.
Approximately, 3350 rooted plants could be generated in 10 mo. after eight subcultures, from one shoot with a shoot apex and
four nodes. 相似文献
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P. S. Sha Valli Khan E. Prakash K. R. Rao 《In vitro cellular & developmental biology. Plant》2002,38(2):186-190
Summary A protocol has been developed for plantlet regeneration from seed callus of Bixa orellana L. Seeds demonstrated a high percentage of callus induction (63±7.3%) and a high yield (356±14.7 mg per seed) of white friable
callus on Murashige and Skoog (MS) medium containing 5.0 μM l-naphthaleneacetic acid (NAA) and 2.5μM N
6-benzyladenine (BA) within 6 wk of culture in the dark. Callus induction frequency was greater under 24h dark as compared
to 16h light/8h dark photoperiod or 24h light photoperiod. Increased myo-inositol (MI: 200mgl−1) and addition of ascorbic acid (AA: 200 mgl−1) to the culture medium positively improved callus induction frequency and growth. Shoot differentiation from white friable
seed callus was best using 10.0 μM BA and 5.0 μM NAA, where the highest percentage of calluses forming shools (74.9±4.8%), the highest number of shoots per callus (six or
seven) and the highest shoot-forming index (5.0) were obtained within 6 wk. Shoots elongated to 4 cm within 4 wk of transfer
onto MS medium devoid of growth regulators. Shoots were rooted using half-strength MS medium containing 5.0 μM indole-3-butyric acid (IBA). About 85% of these plants were established in pots containing pure garden soil and organic manure
after 3 wk of hardening. Regenerated plants were morphologically uniform with normal leaf, shape and growth patterns. These
plants are currently being screened for the presence of agronomically useful genetic variants. 相似文献
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