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A major limitation with the available protocols for in vitro regeneration of peanut (Arachis hypogaea L.) is their narrow application to very few select genotypes. Here, we report a protocol that can be applied across a broad spectrum of peanut market types, explant types and geographic regions using thidiazuron (TDZ). The effect of the timing of TDZ application to the culturing of both zygotic embryos and subsequent plantlet explants on MS medium is also reported. An extended use of TDZ and at a higher concentration (30 m/l) resulted in the greatest explant shoot average (approximately 13). However, a limited application of TDZ (10 d) was sufficient to induce shoot formation in peanut. Hypocotyl was the best explant type that induced the greatest shoot average (15) across market types followed by lamina (7.4). Spanish and Valencia were the most efficient market groups that induced shoots across explant types, consistently. 相似文献
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Ning?WuEmail author Kanyand?MatandEmail author Huijuan?Wu Baoming?Li Kayla?Love Brittany?Stoutermire Yanfeng?Wu 《Central European Journal of Biology》2012,7(4):620-625
The accurate prediction of plant pre-mRNA splicing sites has been studied extensively. The rules for plant pre-mRNA splicing
still remain unknown. This study, based on confirmed sequence data, systematically analyzed all expressed genes on Arabidopsis thaliana chromosome IV to quantitatively explore the natural splicing rules. The results indicated that defining Arabidopsis thaliana pre-mRNA splicing sites required a combination of multiple factors including (1) relative conserved consensus sequence at
splicing site; (2) individual nucleotide distribution pattern in 50 nucleotides up- and down-stream regions of splicing site;
(3) quantitative analysis of individual nucleotide distribution by using the formulations concluded from this study. The combination
of all these factors together can bring the accuracy of Arabidopsis thaliana splicing site recognition over 99%. The results provide additional information to the future of plant pre-mRNA splicing research. 相似文献
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ATP-binding cassette transporter G2 (ABCG2) gene encodes a protein that has a wide variety of substrates and is responsible
for the active secretion of clinically and toxicologically important molecules into milk. Although known in many species,
this marks the first time this gene product has been reported in goats. In this study, we cloned and sequenced goat ABCG2
gene complete coding sequence and predicted its putative translated protein structure with implicative functional domains.
One six-transmembrane span on C-terminal region and at least one coiled-coil domain on N-terminal were predicted and compared
primarily with those of other closely related species. In addition, three conserved cysteines (in positions 595, 606, and
611) were determined toward the C-terminal of goat’s ABCG2. Two known functional motifs were identified in goat’s protein
through comparative studies with other species. The goat ABCG2 relative expression profile revealed that the gene expression
was a function of lactation stage and parallel to goat lactation curve. 相似文献
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Han XF Luo J Wu N Matand K Yang BJ Wu HJ Zhang LJ Wang HB 《Molecular biotechnology》2008,38(3):187-193
A lactating goat mammary gland cDNA library was constructed by using a modified commercially available cDNA library construction
kit protocol. The resulting clones were sequenced and functionally analyzed through cross-species genomic comparison to assess
(1) the capacity and functional quality of the constructed library for subsequent research and (2) the efficiency of the procedural
modifications. The study resulted in the construction of a high-quality mammary gland cDNA library, which was characterized
by (1) the total recombinants number of 1.4 × 107 colony-forming units (cfus) that was at least 10 times greater than the number expected from the application of the standard
kit protocol, (2) the recombinants rate of 96%, and (3) the average insert size of 1,082 bp. BLAST analysis of sequenced clones
against GenBank databases determined 55.7% of clone redundancy, 22 known function gene clusters, and 29 novel gene clusters.
The analysis of the primary gene expression profile showed that 59% of the tested clones were genes that coded for milk proteins
while 16% of the clones coded for ribosomal, metabolism, immune response, and translation proteins. The remaining 25% of the
tested clones were described as novel genes. Cross-species comparison showed that 77% of characterized gene clusters were
successfully identified by using resources from other ruminants and unrelated species. This outcome is in consonance with
the common belief that the genomic resources that have been generated across species are potentially powerful tools that could
be used for enhancing the molecular understanding of less genomically studied species, such as goat. 相似文献
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Multiple shoots were induced on Valenciatype peanut (Arachis hypogaea L.) explants cultured in vitro on a nutrient medium supplemented with thidiazuron. Zygotic embryos excised from mature seeds were germinated on Murashige-Skoog nutrient medium, and the resulting plantlets (8 days-old) were used as a source of explants. When cultured on a nutrient medium with increasing levels of thidiazuron (0.5 to 30 mg/l), expiants from various parts of the peanut plant (except the root) produced multiple shoot primordia which subsequently developed into individual shoots. Hypocotyl and cotyledon explants produced shoots in higher numbers than other explants (20 shoots per hypocotyl explant at all thidiazuron concentrations and 15 shoots per cotyledon explant at 30 mg/l). Shoots rooted normally on a basal Murashige-Skoog medium containing charcoal and developed into healthy and fertile plants when planted in soil.Abbreviations TDZ
thidiazuron
- MSO
Murashige and Skoog (1962) basal medium
- BA
6-benzylaminopurine 相似文献
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Subtraction technique has been broadly applied for target gene discovery. However, most current protocols apply relative differential
subtraction and result in great amount clone mixtures of unique and differentially expressed genes. This makes it more difficult
to identify unique or target-orientated expressed genes. In this study, we developed a novel method for subtraction at mRNA
level by integrating magnetic particle technology into driver preparation and tester–driver hybridization to facilitate uniquely
expressed gene discovery between peanut immature pod and leaf through a single round subtraction. The resulting target clones
were further validated through polymerase chain reaction screening using peanut immature pod and leaf cDNA libraries as templates.
This study has resulted in identifying several genes expressed uniquely in immature peanut pod. These target genes can be
used for future peanut functional genome and genetic engineering research. 相似文献
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