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Marker Assisted Selection-A New Paradigm in Plant Breeding
Kshirod K. Jena,Huhn Pal Moon,David J. Mackill 한국육종학회 2003 한국육종학회지 Vol.35 No.3
The production and productivity of major crop plants have reached their plateu during the past decade because of adop-tion of green revolution technologies. However, further increase in production of cereals with improved cereal quality is imperativeto fe
Effectiveness of marker-assisted backcrossing breeding for biotic resistance in rice
Jung-Pil Suh,Kshirod K. Jena,Young-Chan Cho,Ji-Ung Jeung,Yong-Jae Won,Im-Soo Choi,Jeom-Ho Lee,Myeong-Ki Kim,Chung-Kon Kim 한국육종학회 2013 한국육종학회 심포지엄 Vol.2013 No.07
The transfer of a biotic resistance gene from indica rice cultivars into japonica cultivars by conventional breeding methods often difficult due to high sterility of the progenies, poor plant type, and linkage drag. Molecular markers provide opportunities to map resistance genes and accelerate the application of marker-assisted backcross(MAB) breeding through the precise transfer of target genomic regions into the recurrent parent. The basis of MAB breeding is to transfer a specific gene/allele of the donor parent into the recurrent parent genome while selecting against donor introgressions across the rest of the genome. The effectiveness of MAB breeding depends on the availability of closely linked DNA markers for the target locus, the size of the population, the number of backcrosses and the position and number of markers for background selection. We have successfully developed Bph18 version of the commercially cultivated japonica elite cultivar by using MAB and incorporating the resistance gene Bph18 that conferred enhanced resistance to BPH. MAB breeding provides a new opportunity for the selective transfer of biotic resistance genes into elite indica rice cultivars devoid of linkage drag. In additon, molecular markers precisely estimate the introgression of chromosome segments from donor parents and can speed up the recipient genome recovery via background selection.
A simple, rapid, and high-throughput DNA extraction method for PCR analysis from rice plants
Sung-Ryul Kim,Gynheung An,Kshirod K Jena 한국육종학회 2013 한국육종학회 심포지엄 Vol.2013 No.07
Polymerase chain reaction (PCR) is highly utilized for QTL analysis, positional cloning of valuable genes, and molecular breeding in crop science. Usually those experiments handle DNA samples of many genotypes (up to several thousands). However, many DNA extraction protocols require longer time using harmful chemicals such as chloroform, phenol, and liquid nitrogen. Here, we introduce a new DNA extraction method for PCR with agarose/PAGE analysis from a diversity panel of rice genotypes identified with yield enhancing traits. This protocol consists of four steps including injection of extraction buffer (20 mM Tris-HCl pH9.5, 200 mM KCl, 2 mM EDTA) into the tubes containing leaf tissues and steel balls, and crushing tissues using Geno-Grinder without liquid nitrogen, sample incubation at 65°C, and then centrifugation for removing cell debris. After centrifugation the crude extracts directly used as template DNA for PCR. Through this protocol we could complete F1 hybridity test from approximately 2,100 plants that come from 96 cross combinations with 13 SSR markers. In addition, we tested the DNA quality by PCR amplification of high GC-rich region and large target size (-2kb). From these results our DNA extraction method produces enough DNA quality for PCR and is suitable for large scale molecular analysis from rice plants.
Kasturi Pusty,Kshirod K. Dash,Ajita Tiwari,V. M. BALASUBRAMANIAM 한국식품과학회 2023 Food Science and Biotechnology Vol.32 No.14
In the present study encapsulation of ultrasound assisted red cabbage extract was carried out using four different carrier agents such as maltodextrin, gum arbic, xanthan gum, and gellan gum. Among the four hydrocolloids investigated, maltodextrin was found to have the least destructive effect on anthocyanin content (14.87 mg C3G/g dw), TPC (54.51 ± 0.09 mg GAE/g dw), TFC (19.82 Mg RE/g dw) and antioxidant activity (74.15%) upon freeze–drying. Subsequently a storage study was conducted using maltodextrin as carrier agent at 25–50 °C. The Clausius–Clapeyron equation was used to evaluate the net isosteric heat (qst) of water adsorption. The differential entropy (ΔS) and qst decreased from 82.298 to 38.628 J/mol, and 27.518 kJ/mol to 12.505 kJ/mol, respectively as the moisture content increased from 2 to 14%. The value of isokinetic energy and Gibb’s free energy were found to be 364.88 and − 1.596 kJ/mol for freeze dried red cabbage.
( Sung Ryul Kim ),( Jungil Yang ),( Gynheung An ),( Kshirod K Jena ) 한국육종학회 2016 Plant Breeding and Biotechnology Vol.4 No.1
Preparation of DNA is cumbersome especially in the case of large numbers of plant samples. Several simple plant DNA preparation methods have been developed for use in conjunction with polymerase chain reaction (PCR) analysis. However, those methods have not been adopted widely for rice molecular analysis. We present a new, simple, and inexpensive method using tris-phosphate (TPE) ethylenediaminetetraacetic acid (EDTA) buffer (100 mM tris-HCl pH9.5, 1 M KCl, 10 mM EDTA pH 8.0) without phenol-chloroform extraction and DNA precipitation steps. The method consists of five steps: leaf tissue grinding, incubating in TPE buffer at 65oC for 20 to 90 minutes, diluting extracts with water, centrifuging to sediment tissue debris, and transferring the supernatant for direct use in PCR or storage. Agarose gel analysis of the crude extracts indicated that the method produced intact genomic DNA (gDNA) from young and old leaves of both young seedlings and mature plants. Leaf sample size (0.5 to 8.0 cm long) for DNA preparation was less sensitive to PCR than the previous methods. DNA quality was tested through PCR amplification of various GC content regions and product sizes, and we obtained bands from all samples, indicating that the method produced suitable DNA quality for PCR. gDNAs were stable for longer than eight months at 4oC. This protocol enabled one person to handle several hundred samples in a day and was tested through various PCR-gel analyses such as genotyping of rice T-DNA mutant lines, positional cloning of rice mutant, and high throughput marker-assisted breeding using allele-specific SNP/Indel markers.
G. V. S. Bhagya Raj,Kshirod K. Dash 한국식품과학회 2023 Food Science and Biotechnology Vol.32 No.6
The pearl millet based functional pasta was formulated by incorporating freeze dried dragon fruit pulp powder and 2% (w/w) microcapsule containing dragon fruit peel extract. The control pasta consisted of 100% pearl millet flour. The other four functional pasta samples consisted of pearl millet and freeze-dried dragon fruit pulp powder (DFP) in the ratio of 95:5, 90:10, 85:15, and 80:20 (w/w), respectively. The inclusion of dragon fruit powder enhanced the swelling index, water absorption index, color, and functional properties of the pasta. The total phenolic content (0.24–0.43 mg GAE/100 g d.w.), antioxidant activity (17.76–30.67%), and betacyanin content (0.149–0.152 mg/g d.w.) of the pasta was increased with the increase of dragon fruit pulp level in the formulation. The release kinetics of phenolic compounds into the simulated gastric juice was modeled using Higuchi and Peppas- Sahlin models. Out of these two models Peppas- Sahlin model (R2>0.980and RMSE<1.527) found to predict the release of phenolics into simulated gastric juice with respect to time of release when compared with Higuchi model (R2>0.964and RMSE<6.126). The onset of transition temperature and enthalpy of gelatinization of pasta samples was found to be in the range of 66.321–74.681 °C and increased with the increase of dragon fruit level in the formulation.
Effect of intermittent microwave convective drying on physicochemical properties of dragon fruit
G. V. S. Bhagya Raj,Kshirod K. Dash 한국식품과학회 2022 Food Science and Biotechnology Vol.31 No.5
The study was carried out to investigate the effect of Intermittent microwave convective drying (IMCD) on the overall quality of dried dragon fruit in terms of total phenolic content, color change, and rehydration ratio. Three levels of microwave power (200–600 W) and a temperature of 60 °C for hot air were applied alternately throughout the process with three levels of pulse ratio such as 1:10, 1:20, and 1:40, respectively. The total phenolic content of the dragon fruit slice obtained by IMCD was ranged between 5.750 and 6.575 mg GAE/g dry weight. Within the experimental range of process variables under IMCD conditions, the drying efficiency, color change, and rehydration ratio of the dried dragon fruit slices were 15.287–51.930%, 18.643–24.847, and 1.908–3.239, respectively. The Weibull model scale (α) parameter was found to vary between 27.512-498.174, while the shape (β) parameter was found to vary between 0.769−0.851. The Weibull model parameters were shown to decrease with increasing microwave power at constant pulse ratio. The IMCD method produced a dried dragon fruit slices with reduced color changes and higher total phenolic content and rehydration ratio values. This investigation would contribute to the development of effective drying techniques for increased food quality and product consistency in the drying of diverse fruits and vegetables.