, and after storage ( Duan et al., 2017 ; Leisso et al., 2016 ; Nham et al., 2015 ). Global-scale gene expression analysis afforded by second-generation sequencing of mRNA allows the activity of all genes to be monitored simultaneously. This powerful
Heidi Hargarten, Sumyya Waliullah, Lee Kalcsits, and Loren A. Honaas
Cai-Hong Jia, Ju-Hua Liu, Zhi-Qiang Jin, Qiu-Ju Deng, Jian-Bin Zhang, and Bi-Yu Xu
analyzed by BLAST (< http://ncbi.nlm.nih.gov/blast >). Quantitative real-time RT-PCR analysis of MaMDH expression. For real-time quantitative reverse transcription (RT)–PCR, total RNA was extracted from the roots, rhizomes, leaves, flowers, and fruit
Yingmei Gao, Jingkang Hu, Tingting Zhao, Xiangyang Xu, Jingbin Jiang, and Jingfu Li
was adopted to analyze relative gene expression. RNA expression levels relative to the actin gene were calculated as 2 −△△CT values according to a previous analysis ( Pfaffl, 2001 ). The primers used for qRT-PCR were designed according to the genomic
Zhigang Ouyang, Huihui Duan, Lanfang Mi, Wei Hu, Jianmei Chen, Xingtao Li, and Balian Zhong
(LSM 510 Meta confocal microscope; Zeiss, Oberkochen, Germany). RNA sequencing. Transcriptome sequencing (RNA-Seq) analysis was used to study the expression of CitYTH genes during different fruit development stages. Transcriptome sequencing and assembly
Jingkang Hu, Yingmei Gao, Tingting Zhao, Jingfu Li, Meini Yao, and Xiangyang Xu
with tandem duplications are shown in different colors, and gene clusters were marked by black lines. Expression analysis of ZF-HD genes in different tissues. Expression patterns of 15 ZF-HD genes in six tissues of tomato ‘Moneymaker’ plants were
Wanyu Xu, Chen Chen, Ningning Gou, Mengzhen Huang, Tana Wuyun, Gaopu Zhu, Han Zhao, Huimin Liu, and Lin Wang
to synonymous substitutions (Ka/Ks) value, where Ka/Ks <1 indicates a negative selection, Ka/Ks = 1 indicates a neutral selection, and Ka/Ks >1 indicates a positive selection ( Gong et al., 2019 ). Expression analysis using RNA-sequencing data
C.S. Prakash, O. Zheng, and A. Porobodessai
Stable, transgenic, sweetpotato plants have been developed using an improved somatic embryogenesis consisting of l) stage I—explants incubated in darkness for 14 days on MS medium with 2,4D (2.5 mg·liter–1) and 6-BAP (0.25 mg·liter–1) and 2) stage II—culture in light for 14 to 28 days on MS medium with ABA (2.5 mg·liter–1). Petiole or leaf explants of the genotype PI318846-3 were co-cultivated with Agrobacterium tumefaciens EHA 101 containing gusA::nptII fusion gene. Transgenic somatic embryos were selected on a kanamycin medium (100 mg·liter–1). The PCR analysis of the transgenic sweetpotato plants showed the presence of foreign genes in the sweetpotato genome. About 100 transgenic plants are being maintained under laboratory and greenhouse conditions. All the transgenic plants showed a strong expression of gusA gene in the histochemical GUS assay but showed quantitative differences in the chemiluminescent assay. The CaMV35S promoter shows a differential expression because there was some degree of tissue- and organ-specificity in the gusA expression. All transgenic plants appear normal with no phenotypic aberrations and are being tested for productivity traits.
Xianqin Qiu, Hongying Jian, Qigang Wang, Kaixue Tang, and Manzhu Bao
are therefore candidate Mlo orthologs. Gene expression analysis has become increasingly important in many fields of biological research. Understanding patterns of expressed genes is expected to provide insight into complex regulatory networks and
Ying Jia, Dianren Xia, and E.S. Louzada
A cDNA coding for a putative terpene synthase (Grtps) was isolated from `Rio Red' grapefruit (Citrus paradisi Macf.) mature fruit by differential display RT-PCR and the corresponding full-length cDNA and genomic clone were subsequently obtained. The isolated cDNA clone was 1644 bp in length encoding a protein of 548 amino acids with a predicted molecular mass of 64 kDa and of pI 5.38. The genomic clone was 3203 bp in length with 6 introns and 7 exons. This Grtps appears to be a sesquiterpene synthase based on molecular weight, genomic organization, and similarity with the other terpene synthases. Both RT-PCR and Northern blot expression analysis indicated that Grtps is not expressed in immature fruits, roots, or leaves, but only in mature fruits. Southern blot analysis of genomic DNA demonstrated that Grtps is one of the members in the family of terpene synthases.
Adriana Robbins, Ying Jia, and Eliezer Louzada*
In Texas, the freezes of 1951 and 1962 together killed 125,000 acres of citrus trees and the freeze of 1983 killed 40,000 acres. The low temperature is one of the most important abiotic stresses to be understood and manipulated molecularly. Cold hardiness is found in the deciduous citrus relative, trifoliate orange, which can withstand temperatures as low as -26 °C when it is cold acclimated. Exposure of the cold hardy trifoliate orange plants to temperature from 28 °C to -5 °C enabled us to isolate and characterize one novel citrus low temperature gene (clt) with two transcripts, called clt-a and clt-b from leaves and twigs. Clt-a was produced when plants were subjected to low temperatures (starting at 10 °C), while cltb was constitutively expressed. Both clt-a and clt-b have the same open reading frame of 165 nucleotides and encodes a small protein of 54 amino acid. However, clt-a has an additional 98 bp nucleotides at the 3'-untranslated region (UTR), which is absent in clt-b. Expression analysis using relative quantitative RT-PCR demonstrated that clt-a is expressed exclusively at low temperatures, while clt-b is expressed constitutively (expression verified from 2 °C to -5 °C). In the process of deacclimation from -1 °C to 28 °C, the clt-a transcript degraded dramatically after 2 °C and was completely absent at 28 °C, while the clt-b transcript remain stable. When the acclimated plant was taken from -1 °C to room temperature, the clt-a gene degraded within 2 hours. Moreover, when acclimated plant was continuously exposed at -1 °C for 20 days, both transcripts clt-a and clt-b remained stable. Involvement of alternative splicing in transcript stability will be discussed.