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Molecular cloning and characterization of an ethylene receptor gene inpotato (Solanum tuberosum L.)孫嘉華, Sun, Ka-wah. January 2000 (has links)
published_or_final_version / Botany / Doctoral / Doctor of Philosophy
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Molecular phylogeny and genetic diversity of sweetpotato (Ipomoea batatas) and its wild relatives黃俊潮, Huang, Junchao. January 2000 (has links)
published_or_final_version / Zoology / Doctoral / Doctor of Philosophy
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Gene expression analysis of the proteinase inhibitor gene PiA of potato (Solanum tuberosum) by reverse transcription polymerase chain reaction (RT-PCR)Zhang, Tieling, 1968- January 2002 (has links)
In this study the gene expression pattern of a novel proteinase inhibitor II (PI-II) gene, PiA, in potato (Solanum tuberosum ), was analyzed using RT-PCR. The PiA gene was found to have enhanced expression in expanding tubers of a greenhouse-grown diploid clone (DC), different stages of 'Russet Burbank' tubers and expanding microtubers of in vitro-grown 'Russet Burbank' and 'Bintje'. PiA mRNA was more abundant in the tuber vascular ring and perimedullary zone areas than in other tuber tissues of DC. The PiA gene was also expressed in flower buds/flowers, shoot apices, leaf blades, stems and roots of 'Russet Burbank'. The PiA mRNA levels did not show significant change after IAA treatment in detached leaf blades and tubers of 'Russet Burbank'. Results suggest that the PiA gene is constitutively expressed and could be involved in tuber developmental processes. To the best of our knowledge, PiA is the first reported PI-II gene expressed in most tissues/organs of potato plants.
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Gene expression analysis of the proteinase inhibitor gene PiA of potato (Solanum tuberosum) by reverse transcription polymerase chain reaction (RT-PCR)Zhang, Tieling, 1968- January 2002 (has links)
No description available.
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Dormancy and germination of true potato (Solanum tuberosum L.) seeds : characterization of endo-β-mannanase genesMonteros, Alvaro R. 06 December 2002 (has links)
True potato (Solanum tuberosum) seed (TPS) is used for preservation of
variable genetic lines of wild and cultivated potatoes (Hawkes et al., 2000) and for
propagation of food crops in some developing countries. TPS has advantages over
seed potato tubers in storage and transportation and favors lower virus infection
levels in fields. However, TPS has thermodormancy and will not readily germinate
at 25°C and above (D'Antonio and McHale, 1988; Pallais, 1995a, b; Alvarado et
al., 2000). TPS can be extremely unreliable when planted directly in fields due to
poor emergence related to diseases and soil crusting.
Germination tests were conducted with two lots of TPS derived from cvs.
EB-8109 and All Blue, respectively, to study dormancy mechanisms. Seeds were
germinated under four temperature regimes (10°C, 15°C, 20°C and 25°C). The two
lots showed distinctly different germination characteristics. EB-8109 seeds showed
only thermodormancy whereas All Blue seeds showed very deep dormancy.
A carotenoid synthesis inhibitor, fluridone, which blocks abscisic acid
(ABA) synthesis, effectively broke thermodormancy in EB-8109 TPS but did not
break primary dormancy in All Blue seeds. Additional treatments, including pre-chilling
and hormonal regimes, also failed to break All Blue deep dormancy. When
the micropylar region of the endosperm (endosperm cap) was removed from seeds
of both seed lots, radicle elongation was observed, suggesting that mechanical
resistance from the endosperm cap restrains radicle protrusion, and that weakening
of the endosperm cap is requisite for TPS germination.
Endo-β-mannanase expression was measured to help characterize
mechanism underlying the weakening of endosperm cap tissues. This enzyme is
thought to permit radicle protrusion by degrading cell walls thereby weakening the
tissues of the endosperm cap (Groot et al., 1988). The coding region of
germination-specific mannanase was isolated from the potato genome by use of
polymerase chain reaction (PCR) with primers specifically designed for the tomato
germination-specific mannanase gene (LeMAN2, Nonogaki et al., 2000). The
cDNA of the TPS mannanase was identical to that of LeMAN2. The expression of
mannanase mRNA was detected in the endosperm cap of germinating TPS after 72
h of imbibition at 15°C, while no expression was detected at 25°C (thermodormant
condition). Fluridone induced mannanase expression in the micropylar region of
the endospem at 25°C. Thus, there was a correlation between induction of
mannanase and dormancy breakage.
A major increase in TPS post-germinative endo-β-mannanase activity was
detected by use of gel diffusion assay. Two isoforms of mannanases were detected
in the protein extracts of germinated TPS by activity staining of native
polyacrylamide gel electrophoresis. The post-germinative mannanase was detected
in the whole endosperm of germinated TPS by using tissue printing with the
LeMAN1 (Bewley et al., 1997) RNA probe. These results suggest that, as with
tomato, TPS also expresses post-germinative mannanase activity.
The promoter region of a new tomato mannanase was isolated during this
research. This promoter was shown to be involved in anther-specific expression of
mannanase. / Graduation date: 2003
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TUBER RUSSET PHENOCOPIES IN POTATO (SOLANUM TUBEROSUM L.) INDUCED BY MEFLUIDIDE.Bidja Mankono, J. Emmanuel. January 1982 (has links)
No description available.
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Another culture of Solanum genotypes.Liebenberg, Denise. January 1995 (has links)
Being the third most cultivated crop in South Africa, potatoes are of great economic
importance. As potatoes originated from cooler areas in the world, they do not easily
adapt to South African conditions. The main objective of potato breeding is, therefore,
to extend the crop's limited genetic base. Progress in crop improvement is slow due
to dominance, segregation and other factors caused by the tetraploid character of
cultivated potatoes. A new breeding program for rapid progress has been initiated at
the Vegetable and Ornamental Plant Institute, Roodeplaat, South Africa, which
comprises the combination of conventional and unconventional breeding techniques.
The program is based on the reduction of the ploidy level from the tetraploid to the
dihaploid level to facilitate crossings with diploid wild species. Anther culture is the
preferred technique for the rapid reduction of the ploidy level and has been
successfully applied on different members of the Solanaceae. Cultivated potato,
Solanum tuberosum is, however, an important exception.
In this study various potato genotypes (tetraploid cultivars, dihaploid
breeding lines and a diploid wild species) were used in experiments concerning
microtechniques, alternative culture methods and medium manipulation. The main
objectives were to evaluate and compare the androgenetic ability of the various
genotypes used and to try and identify the factors limiting their in vitro response.
Regarding microtechnique, the study focussed on the investigation of the
frequency of androgenesis - as a function of plant age - and the determination of
defined flower bud lenqths representative of the correct microspore developmental
stage for optimal androgenetic response. Combined with an extensive histological
study on the microspore development within anthers, from the time of flower selection,
after a cold-pretreatment and at various time-intervals during the culture period of 42
days, the following conclusions were reached: In vitro androgenetic response proved
optimal when flowers of responsive genotypes were selected during the first seven to
21 days of the flowering period. Both microspore derived embryoid- and callus
development were visible within responsive anthers after a culture period of only seven
days. The flower bud length required for anthers to be in the optimal stage of
microspore development, e.g. the uninucleate stage, varied between the different
genotypes but could readily be determined with the DAPI (4,6-diamidino-2-
phenylindole) technique. It was also concluded that anthers of the tetraploid cultivar
Atzimba should be selected later, between the late-uninucleate and the early-binucleate
developmental stages. This suggested a limited selection period for Atzimba anthers,
as starch depositioning - which prevent embryogenesis - occurs within anthers during the binucleate stage. Histologically, Atzimba showed limited embryoid development
with no embryoid release, while the diploid wild species, S. canasense, proved
androgenetically unresponsive.
Alternative culture methods were applied to study the effect of different culture
phases (liquid, double layered and agar solidified) and anther orientations (lateral,
dorsal and ventral) on the androgenetic response of the potato genotypes used.
Liquid cultures, based on the so-called shed-pollen technique, enhanced the
androgenetic response of the tetraploid cultivar Atzimba. Optimal embryogenesis was
obtained for responsive breeding line 87.2002/3 with the utilization of agar solidified
media, with maximal response when anthers were cultured in the lateral orientation.
No response was observed from S. canasense.
The effect of medium manipulation on the androgenetic response of the three
genotypes was investigated. The utilization of various combinations of different
concentrations of indole-3-acetic acid (1M) and benzyladenine (BA), the alteration of
the initial time of incubation of anthers on the initiation media and the use of media
without growth regulators compared to that containing gibberellic acid (GA[3]), were
investigated. BA had to be present in the initiation media and had a major, though not
exclusive, effect on embryogenesis compared to 1M. The optimal BA concentration
varied between the two trials. IAA also had an increasing effect on anther response,
both in the absence of BA and, especially, in addition with relatively high BA
concentrations. In this experiment, only breeding line 87.2002/3 responded. The
initial culture of anthers, during the first seven to 21 days of the culture period, on
media containing growth regulators proved essential for microspore derived embryoid
production in the tetraploid cultivar Atzimba. As these growth regulators are
metabolized in the culture media, the regular transfer at shorter, two-weekly intervals
to media containing metabolically active substances, proved important. GA[3] had no
enhancing-effect on embryogenesis in any of the three tetraploid cultivars.
The results obtained in this study suggest that the first 21 days is the critical
stage in the anther culture period in terms of the optimal time for flower selection,
embryoid induction and the increase in embryogenetic response due to growth
regulator influence. It is important to pre-determine the developmental stage when
most microspores were in the uninucleate stage of development and to correlate this
stage with a specific flower bud length. This would assure maximum response of
those genotypes amenable to anther culture. It also implies a more practical and
economical starting pOint to anther culture experiments. Following the determination
of microspore developmental stage and pollen fertility, flowers should be selected from
the donor plants only during the first three weeks of the flowering period. The composition of the nutrient media used for potato anther cultures were sufficient with
respect to growth regulators. The growth regulators SA, IAA and the amines
glutamine and asparagine had to be present in the initiation media, especially during
the first three weeks of the culture period. As microspore development within anyone
anther was found to be asynchronous, the regular transfer of anthers to fresh media
is recommended to assure proper development of all microspores. The use of a
slightly higher IAA concentration could be considered, but care should be taken as
too-high concentrations would induce callus production. Microspore derived embryoid
production is preferred, as the ploidy level of callus derived plantlets normally varies
and somaclonal variation can occur. Liquid media should be considered for anther
culture of tetraploid genotypes, while embryoid production can be increased by
culturing the anthers of responsive genotypes on agar solidified media on the lateral
orientation. Finally, the diploid wild species S. canasense seemed androgenetically
unresponsive, or the media and culture conditions used did not satisfy the specific
requirements of this genotype. Androgenetic amenability should first be transferred
by means of interspecific crossings with a responsive dihaploid genotype, such as the
breeding line 87.2002/3. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 1995.
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Evaluation of salt tolerance in potato (Solanum spp.)Khrais, Tala January 1996 (has links)
This research was carried out to identify salt tolerant potato genotypes in vitro among 131 tetraploid potato cultivars (Solanum tuberosum), 9 diploid simple hybrid clones (4 clones of S. chacoense $ times$ S. tuberosum, 4 clones of S. phureja/S. stenotomum $ times$ S. tuberosum, and 1 clone of S. tuberosum $ times$ S. tuberosum), 1 primitive cultivated diploid S. phureja/S. stenotomum accession, 12 tetraploid complex hybrids, and 13 diploid S. chacoense accessions. Four levels of NaCl (0, 40, 80, and 120 mM) were used. The cultivars, and the simple and complex hybrids were tested for salt tolerance at the vegetative stage in the nodal cutting bioassay. The thirteen S. chacoense accessions were tested for salt tolerance at the germination and early seedling growth stage, in a seedling bioassay. Eleven of these S. chacoense accessions were further tested at the vegetative stage, in the nodal cutting bioassay. There was a progressive decline in the morphological parameters measured, with increased salt levels, in the nodal cutting bioassay. The parameters were used collectively in ranking the different genotypes, averaged over three NaCl levels (40, 80, and 120 mM). Twenty potato cultivars, two clones of the simple hybrid S. chacoense $ times$ S. tuberosum, and one complex hybrid were all considered salt tolerant at the vegetative stage. Ranking of seven S. chacoense accessions was similar between early seedling growth and later vegetative stage. Two of these accessions were promising as sources of salt tolerance.
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Evaluation of salt tolerance in potato (Solanum spp.)Khrais, Tala January 1996 (has links)
No description available.
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Derivation, fertility and breeding value of doubled monoploids from the diploid potato species, Solanum phurejaM'Ribu, H. Kabura 01 February 2006 (has links)
Thirty-two monoploids (<i>2n = x = 12</i>) derived by anther culture of ten diploid clones of <i>Solanum phureja</i> were used to generate doubled monoploids through in vitro shoot regeneration. Doubled monoploids were compared to the anther donor and progenitor monoploids for morphological characteristics, and were evaluated for fertility in the greenhouse and progeny performance under field conditions.
Monoploids varied for frequency and earliness of shoot regeneration, number of shoots formed per explant and frequency of chromosome doubling among regenerated shoots. Regeneration was greater when stock plantlets were frequently subcultured (2- or 4-week intervals) and maintained under a 16 h photoperiod, and when explants were incubated at 20°C compared to 25°C. In addition, leaf explants regenerated at higher frequencies than stem explants.
Significant high correlations between monoploids and their doubled monoploids were observed for 14 of 17 characters in the greenhouse. Doubled monoploids were significantly greater than monoploids for 15 characters, indicating a positive effect of increasing gene dosage from monoploid to diploid. The anther donor was not significantly greater than the mean of doubled monoploids for 10 characters; therefore, for specific characters, doubled monoploids without homozygote depression can be obtained.
Doubled monoploids varied for nurnber of days to flower, duration of flowering, abundance of flowers, flower quality, fruit set and seed set; they had lower fruit and seed set than the anther donor. A few clones produced low levels of stainable pollen which had high 2n pollen frequency but did not germinate in vitro. Therefore, they were considered male-sterile for practical purposes. Used as female parents, doubled monoploids were able to transmit the 2n pollen trait to their progenies.
Two of four doubled monoploids exhibited superior general combining ability over the anther donor under field conditions. This demonstrates the potential of passage of a heterozygous genotype through the monoploid sieve. The advantage of the monoploid sieve may be more or less evident depending on the combining ability of the crossing partner and variable performance can be expected among doubled monoploids from an unselected anther donor. The performance of unselected doubled monoploids demonstrates the potential for their utilization in breeding and warrants further research in the area. / Ph. D.
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