DataSheet_1_Multiomics analysis elucidated molecular mechanism of aromatic amino acid biosynthesis in Akebia trifoliata fruit.xlsx

Akebia trifoliata is a novel edible and healthy fruit. Here, we found that this fruit had the highest content of total free amino acids and three aromatic amino acids (AAAs) compared with the other popular fruits, and there was an obvious inverse relationship between AAA and flavonoid levels in various fruit tissues. Multiomics analysis revealed that the evolutionarily strengthened synthetic pathway of all three AAAs, the largely regulating ability conferred by ASP5 in the arogenate pathway and the complementary phenylpyruvate pathway endorsed by ADT of both Phe and Tyr biosynthesis provided reasonable explanations for the high AAA content in the flesh of A. trifoliata fruit. Gene-specific expression could be the main reason for the inverse relationship between AAAs and flavonoids. This study will help us understand the metabolic mechanism of AAAs and to develop A. trifoliata as a fresh fruit crop and medicinal plant by molecular breeding strategies.</p

DataSheet_2_Multiomics analysis elucidated molecular mechanism of aromatic amino acid biosynthesis in Akebia trifoliata fruit.docx

Akebia trifoliata is a novel edible and healthy fruit. Here, we found that this fruit had the highest content of total free amino acids and three aromatic amino acids (AAAs) compared with the other popular fruits, and there was an obvious inverse relationship between AAA and flavonoid levels in various fruit tissues. Multiomics analysis revealed that the evolutionarily strengthened synthetic pathway of all three AAAs, the largely regulating ability conferred by ASP5 in the arogenate pathway and the complementary phenylpyruvate pathway endorsed by ADT of both Phe and Tyr biosynthesis provided reasonable explanations for the high AAA content in the flesh of A. trifoliata fruit. Gene-specific expression could be the main reason for the inverse relationship between AAAs and flavonoids. This study will help us understand the metabolic mechanism of AAAs and to develop A. trifoliata as a fresh fruit crop and medicinal plant by molecular breeding strategies.</p

Expression profiles of the <i>TaLHY</i> gene in circadian rhythm.

<p>The relative expression of <i>TaLHY</i> was normalized to the transcript abundances at 8 a.m. (as 1). The error bars represent the standard deviation among three biological replicates. (units: %).</p

A representative phylogenetic tree of <i>TaLHY</i> and selected LHY genes.

<p>Five LHY sequences of monocotyledon plant species from <i>Brachypodium distachyon</i>, <i>Hordeum vulgare</i>, <i>Oryza Sativa</i>, <i>Setaria italica</i> and <i>Zea mays</i>, and eight LHY sequences of dicotyledon plant species from <i>Solanum lycopersicum</i>, <i>Solanum tuberosum</i>, <i>Arabidopsis</i>, <i>Cicer arietinum</i>, <i>Glycine max</i>, <i>Prunu smume</i>, <i>Vitis vinifera</i> and <i>Cucumis sativus</i> were selected. GeneBank accession numbers are provided after the gene names.</p

Expression profiles of the <i>TaLHY</i> gene in different wheat tissues.

<p>The relative expression of <i>TaLHY</i> was normalized to the transcript abundances in leaves (as 1). The error bars represent the standard deviation among three biological replicates. The asterisks indicate statistically significant variation calculated using Student’s t-test. (*P<0.05; ** P<0.01).</p

The relative expression of <i>TaLHY</i> in wheat Chuannong19 in response to transfection with BSMV:<i>TaLHY</i>.

<p>The relative expression of <i>TaLHY</i> was normalized to the transcript abundances in Mock (as 1). The error bars represent the standard deviation among three biological replicates.</p

<i>TaLHY</i>, a 1R-MYB Transcription Factor, Plays an Important Role in Disease Resistance against Stripe Rust Fungus and Ear Heading in Wheat

<div><p>LHY (late elongated hypocotyl) is an important gene that regulates and controls biological rhythms in plants. Additionally, LHY is highly expressed in the SSH (suppression subtractive hybridization) cDNA library-induced stripe rust pathogen (CYR32) in our previous research. To identify the function of the LHY gene in disease resistance against stripe rust, we used RACE-PCR technology to clone <i>TaLHY</i> in the wheat variety Chuannong19. The cDNA of <i>TaLHY</i> is 3085 bp long with an open reading frame of 1947 bp. <i>TaLHY</i> is speculated to encode a 70.3 kDa protein of 648 amino acids , which has one typical plant MYB-DNA binding domain; additionally, phylogenetic tree shows that <i>TaLHY</i> has the highest homology with LHY of <i>Brachypodium distachyon</i>(<i>BdLHY-like</i>). Quantitative fluorescence PCR indicates that <i>TaLHY</i> has higher expression in the leaf, ear and stem of wheat but lower expression in the root. Infestation of CYR32 can result in up-regulated expression of <i>TaLHY</i>, peaking at 72 h. Using VIGS (virus-induced gene silencing) technology to disease-resistant wheat in the fourth leaf stage, plants with silenced <i>TaLHY</i> cannot complete their heading stage. Through the compatible interaction with the stripe rust physiological race CYR32, Chuannong 19 loses its immune capability toward the stripe rust pathogen, indicating that <i>TaLHY</i> may regulate and participate in the heading of wheat, as well as the defense responses against stripe rust infection.</p></div

Changes in Chlorophyll Content Between CN12 and CN19 Under Shading.

<p>(A), Chlorophyll a (<i>Chl a</i>) content; (B), chlorophyll b (<i>Chl b</i>) content; (C), total chlorophyll content; and (D), <i>Chl a</i>/<i>b</i>. Bars represent the mean ± (SE). Asterisks represent statistically significant differences, as follows: **P≤0.01 and *P≤0.05. Letters represent the probability of multiple comparisons of the means of different genotypes and treatments at each time point, as follows: capital letter, P≤0.01; lowercase letter, P≤0.05. An asterisk in the trend line represents the difference between two adjacent time points for the same genotype and treatment.</p

The growth of wheat plants after gene silencing.

<p>The left two were control Chuannong19 plants, with normal heading and flowering. The right two were <i>TaLHY</i>-silenced plants, unable to enter the heading stage of wheat development.</p

Additional file 1 of Conserved DNA sequence analysis reveals the phylogeography and evolutionary events of Akebia trifoliata in the region across the eastern edge of the Tibetan Plateau and subtropical China

Supplementary Table S1: Locations of populations of Akebia trifoliata sampled, sample sizes(n), the cpDNA haplotype and ITS haplotype frequency, haplotype distribution type, haplotype (gene) diversity (Hd) and nucleotide diversity (π × 10 − 3) of each populatio