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1.
Manoj Kumar Harpal Singh Anoop Kumar Shukla Praveen Chandra Verma Pradhyumna Kumar Singh 《Plant signaling & behavior》2013,8(10)
Embryogenesis in cotton is a difficult task due its genome dependency. We used 3 cotton cultivars (Khandwa-2, G. Cot. 10, and BC-68–2) and Coker-312 as control for regeneration. Efficient somatic embryogenesis was induced in agronomically important Indian cotton cultivars, Khandwa-2 and G. Cot. 10. For callusing in all the cultivars, different media combinations were tried. Embryogenesis was initiated on a hormone-free MS medium (MSB). For embryo maturation and recovery excess of L-glutamine and l-asparagine were used. Khandwa-2 somatic embryos were successfully regenerated into plants. However, no plantlet was obtained in case of G. Cot. 10. Callus induction was also observed in BC-68–2 but there was no embryogenesis observed. The study indicated that the medium and genotype significantly effects embryogenesis. An efficient protocol is described here for regenerating plants via somatic embryogenesis in an elite Indian cotton cultivar Khandwa-2. 相似文献
2.
Plant regeneration via somatic embryogenesis in cotton 总被引:6,自引:0,他引:6
An efficient in vitro plant regeneration system characterized by rapid and continuous production of somatic embryos using leaf and stem explants
of abnormal seedling as an explant have been developed in Gossypium hirsutum L. Embryogenic callus and somatic embryos have been obtained directly from the explants of cotton abnormal seedlings. Plant
growth regulators influenced the induction of cotton somatic embryogenesis. The optimal medium for direct somatic embryogenesis
was modified MS medium supplemented with 0.1 mg l-1 ZT and 2 g l-1 activated carbon. On this medium, an average of 28.0 and 28.1 matured somatic embryos formed from per leaf and stem explants
respectively. The highest frequency of somatic embryogenesis was 100%. The somatic embryos were converted into normal plantlets
when cultured on modified MS medium supplemented with 0.1 mg l-1 ZT. Upon transfer to soil, plants grew well and appeared normal. Plants could be regenerated within 60–80 days. The system
of cotton somatic embryogenesis and plant regeneration described here will facilitate the application of plant tissue culture
and genetic engineering on cotton genetic improvement.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
3.
Summary The diploid cotton species can constitute a valuable gene pool for the more agronomically desirable cultivated tetraploid
cultivars and offer better opportunities to study gene structure and function through gene knockouts. In order to exploit
these advantages, a regeneration system is required to achieve these transformation-based goals. Carbohydrate source and concentration
were evaluated to improve somatic embryo (SE) production and desiccation treatments to improve the conversion efficiency of
SEs to plants in a diploid Gossypium arboreum accession, A2-9 (PI-529712). Improved SE numbers and their subsequent conversion into plantlets was achieved with a Murashige and Skoog
(MS)/sucrose-based medium M2 [0.04M sucrose, 0.3 μM α-naphthaleneacetic acid (NAA)] On this medium, 219 embryos per g initiated, and close to 11% of these embryos germinated
into plantlets. Neither a 5-d desiccation treatment of embryogenic callus previously cultured in liquid medium nor filter
paper insertion improved the numbers of SEs induced or their conversion to plantlets. A 3-d desiccation period resulted in
improved plant regeneration. When immature G. arboreum SEs induced on M1 (0.2M glucose, 2.6 μM NAA, and 0.2 μM kinetin) medium underwent a 3-d desiccation treatment, 49% of these immature SEs were converted to plantlets after a 4-wk
period on M2 medium. These improved results will help to pave the way for future genetic transformation and associated gene structure
and function studies utilizing G. arboreum. These results, in particular the 3-d desiccation treatment, can also be incorporated into regeneration protocols to improve
the regeneration efficiency of other Gossypium species. 相似文献
4.
High Frequency Somatic Embryogenesis in Cotton 总被引:3,自引:1,他引:2
Y. Aydin Z. Ipekci T. Talas-Oğraş A. Zehir K. Bajrovic N. Gozukirmizi 《Biologia Plantarum》2004,48(4):491-495
A highly reproducible system for efficient somatic embryogenesis was developed to regenerate plantlets from cotton (Gossypium hirsutum L.) cultivars (Nazilli M-503 and Nazilli 143). Shoot apices, hypocotyls and nodes of 10-d-old seedlings were used as explants. High frequency (100 %) embryogenic calli was initiated from all tested explants on Murashige and Skoog (1962) (MS) media supplemented with 1 g dm–3 polyvinylpyrrolidone (PVP), 1 mg dm–3 6-benzylaminopurine (BAP), 0.5 mg dm–3 kinetin for Nazilli M-503 and 1 g dm–3 PVP, 2 mg dm–3 BAP, 0.5 mg dm–3 kinetin for Nazilli-143. Globular stage somatic embryos were produced 4 months after transfer to hormone-free MS medium supplemented with 1 g dm–3 PVP. Subsequent subculture of globular embryos every 3 weeks on hormone-free MS medium led to the development of torpedo and cotyledonary stage embryos with the frequency of 75 and 83.2 % from hypocotyl explants of Nazilli M-503 and Nazilli-143, respectively. Afterwards, mature somatic embryos were isolated and cultured on hormone-free MS medium for germination and development into plantlets. The highest germination frequency (42.9 %) for Nazilli M-503 somatic embryos were obtained on hormone-free MS medium after 5 months with hypocotyl explants, whereas germination frequencies of Nazilli-143 embryos from hypocotyl, node and apex explants varied between 22 – 30 %. 相似文献
5.
Immature and mature zygotic embryos of hexaploid, Triticale var. DT-46 formed an embryogenic callus, with subsequent somatic embryo formation upon subculture to MS (Murashige and Skoog, 1962) or N6 (Chu et al., 1975) nutrient medium supplemented with various concentrations (9.0–22.5 M) of 2,4-dichlorophenoxyacetic acid (2,4-D). Of the two types of explants, embryogenic tissue from immature embryos responded at a higher frequency, to form somatic embryos over the callus surface. Leaf-base segment cultured on to 2,4-D-containing medium formed a tissue which did not form somatic embryos and instead differentiated into shoot-buds. N6 medium proved to be more effective than MS in support of somatic embryogenesis or shoot-bud formation. Regeneration of plantlets occurred on 2,4-D-free basal medium. These in vitro-formed plantlets were successfully transferred to soil and set seed. 相似文献
6.
Direct somatic embryogenesis of Frittilaria meleagris L. was induced using leaf base explants excised from in vitro grown shoots. Somatic embryos occurred at the basal part of leaf explants 4 weeks after culture on a Murashige and Skoog
(MS) medium supplemented with various concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) or kinetin (KIN). The highest
number of somatic embryos (SEs) were formed (9.74) from leaf explant on MS medium supplemented with 0.1 mg dm−3 2,4-D after 4 weeks of culture initiation. An initial exposure to a low concentration of KIN in the medium also enhanced
SEs induction. Our observations by light and scanning electron microscopy revealed that SEs originate directly from the epidermal
and subepidermal layers of leaf explant. The developmental stages of somatic embryogenesis from the first unequal cell division
through the meristematic clusters, multi-cellular globular somatic embryos to the fully formed cotyledonary embryos were determined.
After 4 weeks on MS medium without plant growth regulators, SEs developed into bulblets. 相似文献
7.
The cultivated diploid, Gossypium arboreum L., (A genome) is an invaluable genetic resource for improving modern tetraploid cotton (G. hirsutum L. and G. barbadense L.) cultivars. The objective of this research is to select a set of informative and robust microsatellites for studying genetic
relationships among accessions of geographically diverse G. arboreum cultivars. From more than 1,500 previously developed simple sequence repeat (SSR) markers, 115 genomic (BNL) and EST-derived
(MUCS and MUSS) markers were used to evaluate the allelic diversity of a core panel of G. arboreum accessions. These SSR data enabled advanced genome analyses. A set of 25 SSRs were selected based both upon their high level
of informativeness (PIC ≥ 0.50) and the production of clear PCR bands on agarose gels. Subsequently, 96 accessions representing
a wide spectrum of diversity of G. arboreum cultivars were analyzed with these markers. The 25 SSR loci revealed 75 allelic variants (polymorphisms) ranging from 2 to
4 alleles per locus. The Neighborjoining (NJ) method, based on genetic dissimilarities, revealed that cultivars from geographically
adjacent countries tend to cluster together. Outcomes of this research should be useful in decreasing redundancy of effort
and in constructing a core collection of G. arboreum, important for efficient use of this genetic resource in cotton breeding. 相似文献
8.
Summary Somatic embryos could be induced from embryogenic callus originating from mesocotyl as well as leaf-base segments of Paspalum scrobiculatum on Murashige and Skoog (MS) or Chu et al. (N6) medium supplemented with different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D; 4.5, 9.0, 18.0, and 22.5 μM). N6 medium was better than MS, for both explants, for high-frequency somatic embryogenesis. Also, mesocotyl tissues were relatively
more totipotent than leaf-base segments. The somatic embryos ‘germinated’ and formed plantlets on transfer of embryogenic
calluses to hormone-free MS or N6 regeneration medium. Embryogenic cultures could be maintained on low hormone medium which readily regenerated to form plantlets
on hormone-free medium. A higher frequency of plantlet formation occurred on MS than on N6 medium. In vitro-formed plantlets were gradually acclimatized in the culture room and on transfer to soil flowered and set seed. Somatic embryogenesis
and plantlet regeneration from mesocotyl and leaf-base segments are potentially simpler systems than regeneration from ‘embryonic’
explants such as immature embryos and unemerged inflorescences. 相似文献
9.
Roger A. Barthelson Uzma Qaisar David W. Galbraith 《Plant Molecular Biology Reporter》2010,28(2):334-343
Gossypium arboreum is an Old World relative of the more commonly cultivated commercial species Gossypium hirsutum, a newer genetic line formed in the New World. G. arboreum has the important property that it can be cultivated in severely hot, dry climates. The genome of G. arboreum has not been completely sequenced, and annotation for the genome is not extensive. We studied the genome of G. arboreum by using cross-species hybridization studies with genomic microarrays for the more annotated species, Arabidopsis thaliana and Oryza sativa. Approximately 30% of the probes on the A. thaliana and O. sativa microarrays were hybridized effectively by target samples prepared from G. arboreum genomic DNA. Many of genes tentatively identified by hybridization function in various levels of the stress response. Cross-species
hybridization can provide effective clues as to potentially valuable genes that may be present in a less well-studied species
such as G. arboreum. The stress response genes tentatively identified in these studies should provide useful clues for further studies toward
the development of hardier strains of cotton. 相似文献
10.
Genetic diversity and relationships of diploid and tetraploid cottons revealed using AFLP 总被引:17,自引:0,他引:17
A. M. Abdalla O. U. K. Reddy K. M. El-Zik A. E. Pepper 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2001,102(2-3):222-229
Gossypium species (± 49) represent a vast resource of genetic diversity for the improvement of cultivated cotton. To determine intra-
and inter-specific genetic relationships within a diverse collection of Gossypium taxa, we employed 16 AFLP primer combinations on three diploid species, Gossypium herbaceum L. (A1), Gossypium arboreum L. (A2) and Gossypium raimondii Ulbrich (D5), and 26 AD allotetraploid accessions (Gossypium barbadense L. and Gossypium hirsutum L.). A total of 1180 major AFLP bands were observed; 368 of these (31%) were polymorphic. Genetic similarities among all
taxa ranged from 0.21 (between the diploid species G. arboreum and G. raimondii) up to 0.89 (within G. barbadense). Phenetic trees based on genetic similarities (UPGMA, N-J) were consistent with known taxonomic relationships. In some cases,
well-supported phylogenetic relationships, as well as evidence of genetic reticulation, could also be inferred. UPGMA trees
and principal coordinate analysis based on genetic similarity matrices were used to identify genetically distinct cultivars
that are potentially important sources of germplasm for cotton improvement, particularly of fiber quality traits. We show
that AFLP is useful for estimating genetic relationships across a wide range of taxonomic levels, and for analyzing the evolutionary
and historical development of cotton cultivars at the genomic level.
Received: 17 January 2000 / Accepted: 4 May 2000 相似文献
11.
Immature and mature zygotic embryos of Paspalum scrobiculatum L. cv. PSC 1 cultured on MS or N6 nutrient medium supplemented with different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D), formed embryogenic callus. Induction of embryogenic callus and subsequent somatic embryogenesis was possible at a lower concentration of 2,4-D on N6 than MS medium. Immature embryos were highly totipotent, forming somatic embryos at a higher frequency than mature embryos. Addition of amino acids (L-proline or L-tryptophan) to 2,4-D medium resulted in significant enhancement of embryogenesis on culture of mature embryos. Silver nitrate also supported an increased frequency of embryogenesis. Thus it is possible to have high frequency of somatic embryogenesis on culture of mature embryos, which are available in abundance and with ease than immature embryos. The somatic embryos readily germinated and formed plantlets on hormone-free regeneration medium. The regenerated plantlets were successful on transfer to soil and set seed. 相似文献
12.
Summary A protocol was developed for high frequency somatic embryogenesis and plant regeneration from cotyledon and hypocotyl explants
of Eruca sativa. Explants grown on Murashige and Skoog (MS) medium supplemented with 4.52 μM 2,4-D formed embryogenic callus after 4 wk of culture. Secondary somatic embryos were also produced from primary somatic
embryos on MS medium containing 0.56 μM 2,4-D. Somatic embryos developed into mature embryos on MS medium in the presence of 45 gl−1 polyethylene glycol. After desiccation, somatic embryos developed into plantlets by culturing the mature somatic embryos
on 1/2 x MS medium containing 0.24 μM indole-3-butyric acid. 相似文献
13.
Summary Somatic embryogenesis and plant regeneration have been achieved in Nothapodytes foetida, which is known for its rich source of anti-cancer and anti-AIDS alkaloids. Callus cultures were initiated from immature
zygotic embryos cultured on Murashige and Skoog's (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid, 6-benzyladenine
(BA), and kinetin. MS medium devoid of plant growth regulators favored the development of globular somatic embryos that differentiated
further into plantlets. Plantlet regeneration efficiency was effectively increased on MS medium supplemented with BA. Over
90% of the in vitro plantlets survived when transferred to the soil. Alkaloids were detected in different stages of somatic embryos, regenerated
plantlets, and different parts of the 2-yr-old regenerated plants. The somatic embryos contains camptothecin (0.011% dry weight.
DW) and 9-methoxycamptothecin (0.0028% DW). Two-yearold field-grown plants obtained from somatic embryos were analyzed and
contained higher levels of camptothecin (0.20% DW) and 9-methoxycamptothecin. (0.097% DW) accumulated in roots, followed by
stem and leaves. Alkaloids were quantified and identified by TLC and HPLC. 相似文献
14.
Brassinolide (BR), which is the most biologically active brassinosteroid, was used to examine the potential effect of hormone
on cotton somatic embryogenesis. Ten-day-old cotton (Gossypium hirsutum L., cv. Cooker) seedlings were used for explant source and hypocotyls were removed and cultured on MS basal medium with B5 vitamins supplemented with 1 mg/L 6-benzylaminopurine + 0.5 mg/L kinetin for callus induction. After one month proliferating
calli pieces were collected and cultured on MS basal medium containing various concentrations of BR (0.1, 0.5, 1.0 μM) with
their controls. BR treatments were negatively effective on the fresh weight of calli when compared to control. Differential
somatic embryogenesis maturation rates due to BR treatment were observed. Somatic embryogenesis was stimulated especially
for transition to cotyledonary phase at 0.5 mg/L BR. Histological preparations from embryogenic calli and somatic embryos
at different stages of development revealed the spontaneous polyploidisation during early somatic embryogenesis on BR-treated
calli. Present results suggest that BR negatively effected calli growth, however, had a stimulating role in maturation of
somatic embryos. 相似文献
15.
L. O. das Neves S. R. L. Duque J. S. de Almeida P. S. Fevereiro 《Plant cell reports》1999,18(5):398-405
Medicago truncatula ssp Narbonensis and four genotypes of M. truncatula Gaertn cv. Jemalong were tested for their somatic embryogenesis potential using a two-step protocol. In the first step, embryogenic
callus was induced in folioles isolated from shoots grown in vitro and cultured on Murashige and Skoog (MS) medium supplemented
with 2,4-dichlorophenoxyacetic acid and zeatin. In the second step, somatic embryos were allowed to develop from the induced
callus in MS growth-regulator-free medium. Individual somatic embryos were then isolated and transferred again to growth regulator
free medium where they formed secondary somatic embryos in repetitive cycles. Conversion of somatic embryos into plantlets
was achieved by isolating late-torpedo-phase somatic embryos with distinct cotyledons and reculturing them onto MS growth
regulator free medium. The system of repetitive somatic embryogenesis in M. truncatula described here represents a permanent source of embryogenic material that can be used for the genetic modification of this
species.
Received: 7 August 1997 / Revision received: 22 December 1997 / Accepted: 20 January 1998 相似文献
16.
Junaid Aslam Saeed Ahmad Khan Abdul Jaleel Cheruth Abdul Mujib Maheshwar Pershad Sharma Prem Shanker Srivastava 《Saudi Journal of Biological Sciences》2011,18(4):369-380
An efficient somatic embryogenesis system has been established in six date palm (Phoenix dactylifera L.) cultivars (Barhee, Zardai, Khalasah, Muzati, Shishi and Zart). Somatic embryogenesis (SE) was growth regulators and cultivars dependent. Friable embryogenic callus was induced from excised shoot tips on MS medium supplemented with various auxins particularly 2,4-dichlorophenoxyacetic acid (2,4-D, 1.5 mg 1−l). Suspension culture increased embryogenesis potentiality. Only a-naphthaleneacetic acid (NAA, 0.5 mg 1−1) produced somatic embryos in culture. Somatic embryos germinated and converted into plantlets in N6-benzyladenine (BAP, 0.75 mg 1−l) added medium following a treatment with thidiazuron (TDZ, 1.0 mg 1−l) for maturation. Scanning electron microscopy showed early stages of somatic embryo particularly, globular types, and was in masses. Different developing stages of embryogenesis (heart, torpedo and cotyledonary) were observed under histological preparation of embryogenic callus. Biochemical screening at various stages of somatic embryogenesis (embryogenic callus, somatic embryos, matured, germinated embryos and converted plantlets) of date palm cultivars has been conducted and discussed in detail. The result discussed in this paper indicates that somatic embryos were produced in numbers and converted plantlets can be used as a good source of alternative propagation. Genetic modification to the embryo precursor cell may improve the fruit quality and yield further. 相似文献
17.
Laura Y. Solís-Ramos Sara Nahuath-Dzib Antonio Andrade-Torres Felipe Barredo-Pool Tomas González-Estrada Enrique Castaño de la Serna 《Biologia》2010,65(3):504-511
Capsicum chinense is recalcitrant in in vitro morphogenesis. No efficient, reproducible somatic embryogenesis regeneration system exists for this species, impeding regeneration
from transformed cells. An indirect somatic embryogenesis protocol is developed using mature C. chinense zygotic embryo segments (ZES). The ZES cultured in semi-solid Murashige-Skoog (MS) medium supplemented with 8.9 μM naphthaleneacetic
acid, 11.4 μM indoleacetic acid and 8.9 μM 6-benzylaminopurine, developed an embryogenic callus and 8% of the calli developed
somatic embryos. Torpedo-stage somatic embryos were detached from the callus and subcultured in semi-solid MS medium without
growth regulators, producing a 75% conversion rate to plantlets with well-formed root tissue. Histological analysis showed
the developed structures to have no vascular connection with the callus and to be bipolar, confirming that this protocol induced
formation of viable somatic embryos from mature C. chinense ZES. All acclimated plantlets survived under greenhouse conditions. This protocol will facilitate regeneration of genetically
transformed plants using either biolistics or Agrobacterium tumefaciens approach. 相似文献
18.
Kanniah Rajasekaran John W. Pellow 《In vitro cellular & developmental biology. Plant》1997,33(2):88-91
Summary Regeneration of several varieties of soybean [Glycine max (L.) Merrill] by somatic embryogenesis from cultured epicotyls and primary leaves has been demonstrated. Somatic embryogenesis
was induced from epicotyls and primary leaves when cotyledon halves with the intact zygotic embryo axes were cultured on Murashige
and Skoog (MS) medium supplemented with 10 mg 1−1 (45.2 μM) 2,4-D. Stable, continuously proliferating globular embryo cultures (GEC) were established from small groups of somatic embryos
on MS medium supplemented with 20 mg 1−1 (90.5 μM) 2,4-dichlorophenoxyacetic acid (2,4-D). Rapid multiplication of shoot tips from germinating somatic embryos was achieved
on Cheng’s basal medium (CBO) containing 2.5 mg 1−1 (11.3 μM) 6-benzyladenine. Fertile plants were obtained from individual somatic embryos and in vitro propagated adventitious shoot bud cultures. 相似文献
19.
Direct somatic embryogenesis from mature embryos of sandalwood 总被引:7,自引:0,他引:7
Plants were regenerated from mature zygotic embryos of sandalwood (Santalum album L.) through direct somatic embryogenesis. Somatic embryos were formed directly without any intervening callus phase on zygotic embryos plated on Murashige and Skoog (MS) medium containing thidiazuron or benzylaminopurine. Individual somatic embryos were then isolated and transferred to MS medium without cytokinin on which they formed secondary embryos in repetitive cycles with or without the addition of indole acetic acid to the medium. Conversion of somatic embryos into plantlets was achieved by isolating somatic embryos with distinct cotyledons and reculturing them onto half-strength MS medium with GA3 (1.4 M). Recovered plantlets were acclimatised and grown in the greenhouse. This is the first report on in vitro regeneration via direct somatic embryogenesis of sandalwood. 相似文献
20.
María Laura Vidoz Pablo Klusacek Hebe Yolanda Rey Luis Amado Mroginski 《Plant Cell, Tissue and Organ Culture》2006,86(1):111-115
In vitro protocols for plant regeneration of Arachis correntina through both somatic embryogenesis and organogenesis were developed using immature leaves as explants. Morphologically normal somatic embryos were obtained on culture media composed of 20.70 or 41.41 μM picloram (PIC) with the addition of 0.044 μM 6-benzylaminopurine (BA), resulting in a 33 and 24% of conversion into plants, respectively. The source of explants and the developmental stage of the leaves had a marked effect on somatic embryogenesis. The second folded immature leaves from in vitro growing plants were the most responsive producing up to 30% embryogenesis in MS+41.41 μM PIC. Embryos converted into plants after transfer to MS medium devoid of growth regulators and these plants were successfully acclimatised. Adventitious shoots were obtained on culture media supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) or naphthaleneacetic acid (NAA) with or without 0.044 μM BA, achieving plant regeneration in the induction media. The highest percentage of bud formation was obtained on culture medium composed of␣MS+10.74 μM NAA+0.044 μM BA (12.5%). Roots were formed on all culture media tested. Regenerated plants were transferred to pots and grew well under greenhouse conditions. 相似文献