Doubled Haploids generated through anther culture from an elite long duration rice hybrid, CRHR32: Method optimization and molecular characterization

Rout, Prachitara; Naik, Nupur; Ngangkham, Umakanta; Verma, Ram Lakhan; Katara, Jawahar Lal; Singh, O. N.; Samantaray, Sanghamitra

doi:10.5511/plantbiotechnology.16.0719a

Cited by 34 publications

(40 citation statements)

References 36 publications

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“…Additionally, the production of RH requires a three lines system (A, B, and R lines) which is more tedious and requires skilled personnel. These constraints in hybrid rice attract attention to adopt DH technology as there is a possibility to produce filial 1 (F1)-performing high yielding DH lines from the heterotic F1s of rice by fixing hybrid performance exerting the partial to complete dominance in homozygous lines [ 4 , 5 ].…”

Section: Introductionmentioning

confidence: 99%

Anther Culture Efficiency in Quality Hybrid Rice: A Comparison between Hybrid Rice and Its Ratooned Plants

Pattnaik

Dash

Bhuyan

et al. 2020

Plants

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An immense increase in human population along with diminished lands necessitates the increase of rice production since, it serves the human population as a staple food. Though rice hybrids (RH) are showing considerable yield enhancement over inbreds in terms of both quality and quantity, farmers’ adoption of hybrid rice technology has been much slower than expected because of several constraints such as seed cost and quality. Doubled haploid (DH) technology was considered useful for the development of inbred lines from rice hybrids in a single generation. Androgenesis shows its significance in development of DHs in rice which requires an efficient method to establish the production of large population. To start the anther culture, anthers are the main component of androgenesis to be isolated from unopened spikes. However, the duration of spikes availability for anther culture coupled with the segregation of rice hybrids in the next generation requires the main crop be ratooned to reduce the cost of cultivation. Therefore, the efficiency of the androgenic method was tested in main crop using a quality indica rice hybrid, 27P63 and its ratooned ones. The effects of various factors such as cold temperature pre-treatment of boots, treatment duration, and different combination of plant growth regulators (PGR) on callus response along with shoot regeneration were tested for development of DHs from both ratooned and non-ratooned plants. The N6 medium supplemented with 2.0 mg/L 2,4-D (2,4-dichlrophenoxy acetic acid), 0.5 mg/L BAP (6-benzylamino purine), and 30 g/L maltose was found to be most effective for callusing as compared to MS (Murashige and Skoog) medium. The N6 media inducted calli showed maximum response rate for green shoot regeneration in MS media supplemented with 0.5 mg/L NAA (1-napthaleneacetic acid), 0.5 mg/L Kn (Kinetin; 6-furfurylaminopurine), 1.5 mg/L BAP and 30 g/L sucrose after 2 weeks of culture. The pre-treatment of spikes at 10 °C for 2 d followed by a 7th and 8th d were found to be most effective for callusing as well as for regeneration, producing a total of 343 green plants from ratooned and main rice hybrid, 27P63. Morpho-agronomic trait-based assessment of ploidy status revealed 94.46% diploids, 3.49% polyploids, 0.58% mixploids, and 1.45% haploids. Microsatellite markers could authenticate all 324 fertile diploids as true DHs. Though this study shows a reduction in generation of DHs from ratooned plants as compared to the main crop, manipulation of chemical factors could optimize the method to enhance the production of considerable number of DHs. Utilization of ratooned of hybrid rice in androgenesis would save time and cost of cultivation.

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Section: Introductionmentioning

confidence: 99%

Anther Culture Efficiency in Quality Hybrid Rice: A Comparison between Hybrid Rice and Its Ratooned Plants

Pattnaik

Dash

Bhuyan

et al. 2020

Plants

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“…In rice anther culture, the morphological trait that has been used is the measured distance between the nodes of the last two leaves: the flag leaf, and the penultimate leaf [27,29]. In some cases the panicle length at the time of harvest has also been used as a visually identifiable guide [30,31]. The use of a direct cytological marker such as the degree of starch accumulation in microspores has been identified as more accurate than the internode distance and even more convenient than the laborious nuclear staining process to rapidly assess microspore maturity.…”

Section: Pollen Development Stagementioning

confidence: 99%

“…However, these measurements alone cannot be granted sufficiently reliable [54]. When the diploids arisen from anther cultures were analyzed using SSR markers to determine their source of origin, homozygosity has been detected for all in 150 DHs except for one which turned out as a heterozygote [31]. Therefore, microsatellites and other molecular tools such as isozyme analyses and RAPD markers can be utilized to ascertain the homozygosis confirming that the calluses and plantlets have been arisen from gamete itself and not from other somatic tissues.…”

Section: Ploidy-level Determination and Doubled Haploid Productionmentioning

confidence: 99%

Anther Culture as a Supplementary Tool for Rice Breeding

Mayakaduwa¹,

Silva²

2018

Rice Crop - Current Developments

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There is a timely need to harness biotechnology and related tools to support conventional breeding strategies, overcoming the limitations in rice production and improving quantity and quality as well as climatic and disease stress tolerance of the crop. Anther culture allows immediate fixation of homozygosity through diploidization of regenerated haploid plants and therefore serves as an efficient path for inbred line development. Anther culture has been successfully used to hasten the breeding programs in several crop species including rice. However, associated constraints still prevent the realization of its full potential. Even though anther culture technique has been effective for Japonica rice breeding, applicability for Indica rice remains limited mainly due to inherent recalcitrant genetic background. Constraints associated with Indica rice can be identified as early anther necrosis, poor callus induction and proliferation, extremely low green plant regeneration and frequent albinism. Success of androgenesis is determined by factors such as genotype, physiological status of donor plant, pollen development stage at culture, composition and physical status of culture media, culture incubation conditions and anther pretreatments. This chapter has detailed out the scope for improving the applicability of anther culture technique on rice in order to develop it as a supplementary breeding tool.

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“…В ряде работ показана различная плоидность таких регенерантов, полученных из пыльников одной гибридной комбинации или одного генотипа. Среди них встречаются гаплоиды, удвоенные гаплоиды, триплоиды, тетраплоиды, пентаплоиды, анеуплоиды и миксоплоидные растения [7][8][9][10][11][12][13][14], что влечет за собой соответствующую фенотипическую изменчивость регенерантов [7,[12][13][14]. Гаплоидные растения риса характеризуются в три раза большей изменчивостью по содержанию ядерной ДНК, чем удвоенные гаплоиды [15].…”

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Variability of rice haploids obtained in anther culture in vitro

Ilyushko

Илюшко²,

Romashova

2019

Rossiiskaia selskokhoziaistvennaia nauka

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Morphological variability of haploid plants and doubled haploids of rice obtained on one callus line in anther culture in vitro was studied. The work was carried out on rice plants Oryza sativa L. subspecies japonica Kato varietу Cascade. Regenerant plants of one callus line obtained from one rice anther (four in total) were divided into two or three groups of 20-30 plants, depending on the sample size in order of their differentiation on callus and transplantation on the rooting medium. Two callus lines (15.1 and 18.1) formed half of the haploids, half of the doubled haploids, and two other callus lines (5.1 and 7.2) numerous haploids. On callus lines with numerous haploids (5.1 and 7.2), as the regenant number increases, the size of plants decreases (plant height, number of flowers on the main panicle, number of panicles). On the lines 15.1 and 18.1 between groups of haploids and between groups of doubled haploids statistically significant differences absent. In breeding purposes for the induced doubling of the number of chromosomes in haploid regenerants with antitubulin substances such as colchicine, it is advisable to use plants that form on callus among the first. Between haploids of four callus lines and doubled haploids of two callus lines, statistically significant differences (at p=0.001) were revealed using the Hotelling's T2-criterion, calculated for the whole complex of biometric features. Haploids of different lines differed in three or four of them, doubled haploids on three of the five signs (length of panicle, productive bushiness and plant height). Varieties of interest to breeders may be improved by anther culture in vitro.

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Doubled Haploids generated through anther culture from an elite long duration rice hybrid, CRHR32: Method optimization and molecular characterization

Cited by 34 publications

References 36 publications

Anther Culture Efficiency in Quality Hybrid Rice: A Comparison between Hybrid Rice and Its Ratooned Plants

Anther Culture Efficiency in Quality Hybrid Rice: A Comparison between Hybrid Rice and Its Ratooned Plants

Anther Culture as a Supplementary Tool for Rice Breeding

Variability of rice haploids obtained in anther culture in vitro

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