Table_1_Phytohormone biosynthesis and transcriptional analyses provide insight into the main growth stage of male and female cones Pinus koraiensis.xlsx

The cone is a crucial component of the whole life cycle of gymnosperm and an organ for sexual reproduction of gymnosperms. In Pinus koraiensis, the quantity and development process of male and female cones directly influence seed production, which in turn influences the tree’s economic value. There are, however, due to the lack of genetic information and genomic data, the morphological development and molecular mechanism of female and male cones of P. koraiensis have not been analyzed. Long-term phenological observations were used in this study to document the main process of the growth of both male and female cones. Transcriptome sequencing and endogenous hormone levels at three critical developmental stages were then analyzed to identify the regulatory networks that control these stages of cones development. The most significant plant hormones influencing male and female cones growth were discovered to be gibberellin and brassinosteroids, according to measurements of endogenous hormone content. Additionally, transcriptome sequencing allowed the identification of 71,097 and 31,195 DEGs in male and female cones. The synthesis and control of plant hormones during cones growth were discovered via enrichment analysis of key enrichment pathways. FT and other flowering-related genes were discovered in the coexpression network of flower growth development, which contributed to the growth development of male and female cones of P. koraiensis. The findings of this work offer a cutting-edge foundation for understanding reproductive biology and the molecular mechanisms that control the growth development of male and female cones in P. koraiensis.</p

DataSheet_1_Phytohormone biosynthesis and transcriptional analyses provide insight into the main growth stage of male and female cones Pinus koraiensis.docx

The cone is a crucial component of the whole life cycle of gymnosperm and an organ for sexual reproduction of gymnosperms. In Pinus koraiensis, the quantity and development process of male and female cones directly influence seed production, which in turn influences the tree’s economic value. There are, however, due to the lack of genetic information and genomic data, the morphological development and molecular mechanism of female and male cones of P. koraiensis have not been analyzed. Long-term phenological observations were used in this study to document the main process of the growth of both male and female cones. Transcriptome sequencing and endogenous hormone levels at three critical developmental stages were then analyzed to identify the regulatory networks that control these stages of cones development. The most significant plant hormones influencing male and female cones growth were discovered to be gibberellin and brassinosteroids, according to measurements of endogenous hormone content. Additionally, transcriptome sequencing allowed the identification of 71,097 and 31,195 DEGs in male and female cones. The synthesis and control of plant hormones during cones growth were discovered via enrichment analysis of key enrichment pathways. FT and other flowering-related genes were discovered in the coexpression network of flower growth development, which contributed to the growth development of male and female cones of P. koraiensis. The findings of this work offer a cutting-edge foundation for understanding reproductive biology and the molecular mechanisms that control the growth development of male and female cones in P. koraiensis.</p

Turn-final intonation in English

<div><p>The aim of this study was to determine whether transgenic birch (<i>Betula platyphylla</i>) ectopic overexpressing a late embryogenesis abundant (<i>LEA</i>) gene and a basic leucine zipper (<i>bZIP</i>) gene from the salt-tolerant genus <i>Tamarix</i> (salt cedar) show increased tolerance to salt (NaCl) stress. Co-transfer of <i>TaLEA</i> and <i>ThbZIP</i> in birch under the control of two independent CaMV 35S promoters significantly enhanced salt stress. PCR and northern blot analyses indicated that the two genes were ectopically overexpressed in several dual-gene transgenic birch lines. We compared the effects of salt stress among three transgenic birch lines (L-4, L-5, and L-8) and wild type (WT). In all lines, the net photosynthesis values were higher before salt stress treatment than afterwards. After the salt stress treatment, the transgenic lines L-4 and L-8 showed higher values for photosynthetic traits, chlorophyll fluorescence, peroxidase and superoxide dismutase activities, and lower malondialdehyde and Na⁺ contents, compared with those in WT and L-5. These different responses to salt stress suggested that the transcriptional level of the <i>TaLEA</i> and <i>ThbZIP</i> genes differed among the transgenic lines, resulting in a variety of genetic and phenotypic effects. The results of this research can provide a theoretical basis for the genetic engineering of salt-tolerant trees.</p></div

ANOVA analysis of SOD, POD, MDA and Na⁺ Concentration among different lines and time under salt stress.

<p>ANOVA analysis of SOD, POD, MDA and Na⁺ Concentration among different lines and time under salt stress.</p

NaCl stress-tolerance test of transgenic birch ectopic overexpressing <i>TaLEA</i> and <i>ThbZIP</i>.

<p>Stems of transgenic and wild type plants were cultured on WPM medium containing 2 g L^-1 (A), 4 g L^-1 (B) or 6 g L^-1 NaCl (C). Shoots of transgenic and wild type plants were transferred to 1/2 MS root medium containing 4 g L^-1 NaCl (D), or 6 g L^-1 NaCl (E). Photographs were taken 20 d following stress treatment. WT, wild type; L-4, transgenic line 4; L-8, transgenic line 8.</p

Northern blot analysis of <i>TaLEA</i> and <i>ThbZIP</i> expression in transgenic birches.

<p>Total RNA was extracted from the aerial tissues of one-month-old wild type and transgenic lines. The full length <i>TaLEA</i> and <i>ThbZIP</i> genes labeled with DIG-dUTP were used as probes. rRNA, ribosomal RNAs from different samples; <i>TaLEA</i>, target band of <i>TaLEA</i> gene; <i>ThbZIP</i>, target band of <i>ThbZIP</i> gene; L-4, 5, 8, three transgenic lines both containing <i>TaLEA</i> and <i>ThbZIP</i> genes; WT, wild type plantlet, respectively.</p

Average POD, SOD, MDA and Na⁺ content of different lines in different time under salt stress.

<p>Average POD, SOD, MDA and Na⁺ content of different lines in different time under salt stress.</p

Fv/Fm values of different lines after different days salt stress.

<p>Fv/Fm values of different lines after different days salt stress.</p

Map of the T-DNA construct and identification of overexpressing <i>TaLEA</i> and <i>ThbZIP</i> transgenic birches with PCR.

<p>(A) Schematic of the T-DNA region of the binary vector pROKII-<i>TaLEA</i>-<i>ThbZIP</i>. RB, right border; Pnos, nopaline synthase promoter; <i>NptII</i>, kanamycin resistance gene; Tnos, nopaline synthase terminator; P35S, CaMV 35S promoter; <i>Sma</i> I, <i>Sac</i> I, <i>Bam</i>H I, and <i>Kpn</i> I, four different restriction enzyme sites; <i>TaLEA</i>, <i>TaLEA</i> gene; <i>ThbZIP</i>, <i>ThbZIP</i> gene; LB, left border. Agarose gel electrophoresis of PCR products from wild type and transgenic lines with the primer of <i>NptII</i> (B), <i>TaLEA</i> (C), <i>ThbZIP</i> (D). M, DNA marker; PC, positive control; 1–11, eleven Km resistant lines; L-4, 5, 7, 8, 9, five transgenic lines both containing <i>TaLEA</i> and <i>ThbZIP</i> genes; WT, wild type plantlet; H₂O, double-distilled water as negative control.</p

Pn values of WT under different concentrations salt stress.

<p>Pn values of WT under different concentrations salt stress.</p