scholarly journals Regeneration and Agrobacterium-Mediated Transformation of Japonica Rice Varieties Developed for a Cold Region

2018 ◽  
Vol 54 (No. 4) ◽  
pp. 161-167
Author(s):  
Mingfang Feng ◽  
Jing Cang ◽  
Junhong Wang ◽  
Jian Sun ◽  
Jing Yu ◽  
...  

So far, a large number of transformation systems have been established for japonica rice, but only a few have been reported for cold-region varieties. In our study, we established highly efficient tissue culture systems for two cold-region rice cultivars, Dongnong 427 and Longdao 14. Plant growth regulator (PGR) levels were optimized by an orthogonal experimental design. The culture ability, constituted by induction and differentiation rate, served as the detection index of orthogonal experiments. The optimal combinations of PGRs for callus induction and regeneration of Dongnong 427 and Longdao 14 were 1 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) + 2 mg/l 6-benzyladenine (BA) + 4 mg/l kinetin (KIN) + 0.2 mg/l α-naphthaleneacetic acid (NAA) and 1 mg/l 2,4-D + 4 mg/l 6-BA + 4 mg/l KIN + 0.5 mg/l NAA, respectively. Agrobacterium strain EHA 105 containing the plasmid pCAMBIA1301 was used for transformation. The frequency of transient transformation was expressed as the ratio between the number of calli showing GUS expression and the total number of calli kept for staining. The highest transformation efficiency in Dongnong 427 was obtained when calli were immersed in 0.272 OD<sub>600</sub> (optical density determined at 600 nm) for 10 min. While it was best for Longdao 14 calli to be infected with 0.592 OD<sub>600</sub> for 20 min. Infected calli of the two varieties were co-cultivated on two pieces of sterile filter paper moistened with 1 ml liquid co-cultivation medium for three days. The expression of the GUS gene was confirmed by PCR analysis of plants of both varieties.

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Luomiao Yang ◽  
Jingguo Wang ◽  
Zhenghong Han ◽  
Lei Lei ◽  
Hua Long Liu ◽  
...  

Abstract Background Cold stress caused by low temperatures is an important factor restricting rice production. Identification of cold-tolerance genes that can stably express in cold environments is crucial for molecular rice breeding. Results In this study, we employed high-throughput quantitative trait locus sequencing (QTL-seq) analyses in a 460-individual F2:3 mapping population to identify major QTL genomic regions governing cold tolerance at the seedling stage in rice. A novel major QTL (qCTS6) controlling the survival rate (SR) under low-temperature conditions of 9°C/10 days was mapped on the 2.60-Mb interval on chromosome 6. Twenty-seven single-nucleotide polymorphism (SNP) markers were designed for the qCST6 region based on re-sequencing data, and local QTL mapping was conducted using traditional linkage analysis. Eventually, we mapped qCTS6 to a 96.6-kb region containing 13 annotated genes, of which seven predicted genes contained 13 non-synonymous SNP loci. Quantitative reverse transcription PCR analysis revealed that only Os06g0719500, an OsbZIP54 transcription factor, was strongly induced by cold stress. Haplotype analysis confirmed that +376 bp (T>A) in the OsbZIP54 coding region played a key role in regulating cold tolerance in rice. Conclusion We identified OsbZIP54 as a novel regulatory gene associated with rice cold-responsive traits, with its Dongfu-104 allele showing specific cold-induction expression serving as an important molecular variation for rice improvement. This result is expected to further exploration of the genetic mechanism of rice cold tolerance at the seedling stage and improve cold tolerance in rice varieties by marker-assisted selection.


Agronomy ◽  
2020 ◽  
Vol 11 (1) ◽  
pp. 26
Author(s):  
Tao Sun ◽  
Xin Yang ◽  
Sheng Tang ◽  
Kefeng Han ◽  
Ping He ◽  
...  

Nutrient requirements for single-season rice using the quantitative evaluation of the fertility of tropical soils (QUEFTS) model in China have been estimated in a previous study, which involved all the rice varieties; however, it is unclear whether a similar result can be obtained for different rice varieties. In this study, data were collected from field experiments conducted from 2016 to 2019 in Zhejiang Province, China. The dataset was separated into two parts: japonica/indica hybrid rice and japonica rice. To produce 1000 kg of grain, 13.5 kg N, 3.6 kg P, and 20.4 kg K were required in the above-ground plant dry matter for japonica/indica hybrid rice, and the corresponding internal efficiencies (IEs) were 74.0 kg grain per kg N, 279.1 kg grain per kg P, and 49.1 kg grain per kg K. For japonica rice, 17.6 kg N, 4.1 kg P, and 23.0 kg K were required to produce 1000 kg of grain, and the corresponding IEs were 56.8 kg grain per kg N, 244.6 kg grain per kg P, and 43.5 kg grain per kg K. Field validation experiments indicated that the QUEFTS model could be used to estimate nutrient uptake of different rice varieties. We suggest that variety should be taken into consideration when estimating nutrient uptake for rice using the QUEFTS model, which would improve this model.


1984 ◽  
Vol 62 (7) ◽  
pp. 1393-1397 ◽  
Author(s):  
M. D. Zhou ◽  
T. T. Lee

The callus-promoting activity of most commonly known as well as some rarely tested auxins was compared with that of 2,4-dichlorophenoxyacetic acid (2,4-D) for in vitro culture of the excised embryo of spring and winter wheat (Triticum aestivum L.), cv. Chinese Spring and cv. Fredrick. Different auxins in a concentration range from 1 to 50 μM showed widely different activities. Also the two wheat cultivars responded differently to the auxins. When rapid callus formation with limited root growth was used as the basis for comparison, 2-(2-methyl-4-chlorophenoxy)propionic acid (2-MCPP), α-naphthaleneacetic acid, 3,6-dichloro-2-methoxybenzoic acid (dicamba), 4-amino-3,5,6,trichloropicolinic acid (picloram), γ-(2,4-dichlorophenoxy)butyric acid, 2,4,5-trichlorophenoxyacetic acid, and 2,4,5-trichlorophenoxypropionic acid, in the order of effectiveness, were superior to 2,4,-D for callus induction from the embryo of 'Chinese Spring,' although the concentration required was higher than that of 2,4-D. For the winter wheat 'Fredrick,' however, only picloram, dicamba, and 2-MCPP performed as well as 2,4-D. All auxins tested promoted shoot growth; 2-methyl-4-chlorophenoxypropionic acid was most effective for 'Chinese Spring,' whereas picloram was most effective for 'Fredrick.'


Agronomy ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 2253
Author(s):  
Myrish Pacleb ◽  
O-Young Jeong ◽  
Jeom-Sig Lee ◽  
Thelma Padolina ◽  
Rustum Braceros ◽  
...  

Temperate japonica rice is mainly cultivated in temperate regions. Many temperate japonica varieties have a superior grain quality that is preferred in Northeast Asian countries such as Japan, Korea, and China. The changes in consumers’ preferences in Southeast Asia and Western countries has contributed to increasing the demand for temperate japonica. Most temperate japonica varieties developed in temperate regions typically exhibit extra-early flowering under the short-day conditions in the tropics, which usually results in severely reduced yields. Since 1992, we have been developing temperate japonica varieties that can adapt to tropical environments to meet the increasing demand for temperate japonica rice, having released six varieties in the Philippines. Especially, the yield of one of the temperate japonica varieties, Japonica 7, was comparable to the yields of leading indica varieties in the Philippines. Here, we discuss the current breeding initiatives and future plans for the development of tropical-region-bred temperate japonica rice.


2018 ◽  
Vol 14 (1) ◽  
pp. 1
Author(s):  
Joko Prasetiyono ◽  
Nurul Hidayatun ◽  
Tasliah Tasliah

<p>Indonesia is rich in rice genetic resources, however, only a small number has been used in variety improvement programs. This study aimed to determine the genetic diversity of Indonesian rice varieties using 6K SNP markers. The study was conducted at ICABIOGRAD for DNA isolation and IRRI for SNP marker analysis. Genetic materials were 53 rice genotypes consisting of 49 varieties and 4 check genotypes. SNP markers used were 6K loci. Results showed that among the markers analyzed, only 4,606 SNPs (76.77%) were successfully read. The SNP markers covered all twelve rice chromosomes of 945,178.27 bp. The most common allele observed was GG, whereas the least allele was TG. Dendrograms of the 53 rice varieties analyzed with 4,606 SNPs demonstrated several small groups containing genotypic mixtures between indica and japonica rice, and no groups were found to contain firmly indica or japonica type. Structure analysis (K = 2) with value of 0.8 showed that the 53 rice varieties were divided into several groups and each group consisted of 4 japonica, 2 tropical japonica, 46 indica, and 1 aus rice type, respectively. IR64 and Ciherang proved to have an indica genome, while Rojolele has japonica one. Dupa and Hawara Bunar, usually grouped into tropical japonica rice, were classified as indica type, and Hawara Bunar has perfectly 100% indica type. The results of this study indicated that rice classification (indica-japonica) which is usually classified based only on morphological characters, e.g. grain and leaf shapes, is not enough and classification based on SNP markers should be considered for that purpose.</p>


2019 ◽  
Vol 20 (23) ◽  
pp. 5949 ◽  
Author(s):  
Yan Li ◽  
Caihua Dong ◽  
Ming Hu ◽  
Zetao Bai ◽  
Chaobo Tong ◽  
...  

Brassica napus (oilseed rape) is an economically important oil crop worldwide. Sclerotinia stem rot (SSR) caused by Sclerotinia sclerotiorum is a threat to oilseed rape production. Because the flower petals play pivotal roles in the SSR disease cycle, it is useful to express the resistance-related genes specifically in flowers to hinder further infection with S. sclerotiorum. To screen flower-specific promoters, we first analyzed the transcriptome data from 12 different tissues of the B. napus line ZS11. In total, 249 flower-specific candidate genes with high expression in petals were identified, and the expression patterns of 30 candidate genes were verified by quantitative real-time transcription-PCR (qRT-PCR) analysis. Furthermore, two novel flower-specific promoters (FSP046 and FSP061 promoter) were identified, and the tissue specificity and continuous expression in petals were determined in transgenic Arabidopsis thaliana with fusing the promoters to β-glucuronidase (GUS)-reporter gene. GUS staining, transcript expression pattern, and GUS activity analysis indicated that FSP046 and FSP061 promoter were strictly flower-specific promoters, and FSP046 promoter had a stronger activity. The two promoters were further confirmed to be able to direct GUS expression in B. napus flowers using transient expression system. The transcriptome data and the flower-specific promoters screened in the present study will benefit fundamental research for improving the agronomic traits as well as disease and pest control in a tissue-specific manner.


2017 ◽  
Vol 9 (2) ◽  
pp. 27
Author(s):  
Wang Yue ◽  
Sun Fulai ◽  
Gao Qingrong ◽  
Zhang Yanxia ◽  
Wang Nan ◽  
...  

Branched spike wheat is a hexaploid germplasm with branched rachis on its main rachises, and the crucial period for branched rachises occurrence and development is just after the two ridges stage of shoot apex. Natural [indole-3-acetic acid (IAA), indole-3butyric acid (IBA)] and synthetic [(1-naphthaleneacetic acid (NAA), 2,4-Dichlorophenoxyacetic acid (2,4-D)] auxins were applied at this period to investigate the spike traits, seedling growth and photosynthesis related characters and expression of a putative homologue of the LEAFY in branched spike wheat. The four types of experienced auxins induced similar effects on these foresaid characters, although the impact extents were different among the auxins treatments. More branched rachis, spikelets, fertile florets and longer branched rachis were obtained in plants with IAA and IBA at 0.1 mM or NAA and 2,4-D at 1.0mM than those plants with no auxin treated. Auxin treatments also increased fresh and dry mass, photosynthetic pigment and parameters. TFL, a LEAFY-like gene was cloned in branched spike wheat and TFL mRNA expression was quantified using real-time reverse transcriptase-PCR. Application of the auxins accelerated the rise in TFL expression during the periods of branched rachises occurrence and extension. The data supports the hypothesis that auxins play a central role in the regulation branched spike development and TFL might correlate with the development of branched rachises in branched spike wheat.


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