Antisense Lines Research Articles

Folic acid orchestrates root development linking cell elongation with auxin response and acts independently of the TARGET OF RAPAMYCIN signaling in Arabidopsis thaliana

Folic acid is a precursor of tetrahydrofolate (vitamin B9), which is an essential cofactor in most organisms, acting as a carrier for one-carbon units in enzymatic reactions. In this work, we employed pharmacological, genetic and confocal imaging strategies to unravel the signaling mechanism by which folic acid modulates root growth and development. Folic acid supplementation inhibits primary root elongation and induces lateral root formation in a concentration-dependent manner. An analysis of the expression of cell cycle genes pCycD6;1:GFP and CycB1:uidA, and cell expansion Exp7:uidA showed that folic acid promotes cell division but prevented cell elongation, and this correlated with altered expression of auxin-responsive DR5:GFP gene, and PIN1:PIN1:GFP, PIN3:PIN3:GFP, and PIN7:PIN7:GFP auxin transporters at the columella and vasculature of primary roots, whereas mutants defective in auxin signaling (tir1/afb1/afb2 [receptors], slr1 [repressor] and arf7/arf19 [transcription factors]) were less sensitive to folic acid induced primary root shortening and lateral root proliferation. Comparison of growth of WT and TARGET OF RAPAMYCIN (TOR) antisense lines indicates that folic acid acts by an alternative mechanism to this central regulator. Thus, folic acid modulation of root architecture involves auxin and acts independently of the TOR kinase to influence basic cellular programs.

Inhibitory effect of StCYP707A1 gene on tuberization in transgenic potato

Tuber formation of potato is a complex biological process and is regulated by many factors including phytochrome, hormones, transcription factors, RNAs, microRNAs, etc. In our previous study, CYP707A1, an ABA 8′-hydroxylase gene, was down-regulated in the StCOL antisense transgenic potato stolons, but its relation with potato tuberization has not been studied yet. In this study, to investigate the role of this gene in potato tuberization, we cloned a CYP707A1 gene from potato cultivar Desiree and constructed the StCYP707A1 over-expression and anti-sense potato lines by Agrobacterium-mediated transformation. In total, four over-expression transgenic lines and nine anti-sense transgenic lines were confirmed by PCR analysis. Semi-quantitative RT-PCR and qRT-PCR analysis showed that the level of StCYP707A1 transcripts was significantly higher in the over-expression lines (A7 and A9) and lower in the anti-sense lines (F2 and F8), compared to the wild-type control. We further analyzed tuber formation in the transgenic lines and the wild-type control. The result showed that tuber yield per plant and average tuber weight were decreased in A7 and A9 and increased in F2. We also measured the content of ABA and GA3 in transgenic lines. ABA level was reduced in A7 and A9 and increased in F2. Contrariwise, the concentration of GA3 was higher in A7 and A9, and lower in F2 than in wild-type control. These results indicate that StCYP707A1 negatively affects potato tuberization through ABA regulation on gibberellic acid concentration.

Accumulation of phytoene, a colorless carotenoid by inhibition of phytoene desaturase (PDS) gene in Dunaliella salina V-101

The aim of this work was to study the accumulation of phytoene in Dunaliella salina V-101 by down-regulating its phytoene desaturase (PDS) gene expression using RNA interference and Antisense technology. RNAi and antisense constructs were introduced into the Dunaliella cells by Agrobacterium-mediated transformation. Among thirty-two transformants, six showed positive down-regulation of PDS expression with RNAi construct and five positive transformants were obtained using antisense construct. Characterization of PDS suppression was carried out using semi-quantitative RT-PCR and quantitative determination of phytoene as well as other carotenoids by HPLC. Both the RNAi and antisense lines showed a significant decrease in the expression levels of phytoene desaturase and carotenoid content compared to wild type cells. The RNAi line #5 showed maximum Phytoene content (108.34±22.34µg/100mg DCW) compared to other transgenic lines. These phytoene-accumulating phenotypes exhibited slower growth rates and were found to be sensitive to high light conditions.

Alterations in Ascorbic Acid Levels by Transformation of the L-galactono-1,4-lactone Dehydrogenase Gene in Solanum tuberosum

L-galactono-1, 4-lactone dehydrogenase (GLDH) is an important enzyme that catalyzes the last step of the ascorbate biosynthetic pathways in plants. A full-length cDNA clone encoding GLDH was isolated from potato (Solanum tuberosum L. ‘Favorita’) leaf and subcloned into a binary vector, pBI121, to construct sense and antisense recombinant plant expression vectors. The recombinants were introduced into potato via Agrobacterium-mediated transformation, and plants were confirmed as transgenic using PCR and quantitative real-time PCR. Two anti-sense potato lines (G1 and G2) and three sense lines (G3, G4 and G5) were obtained. The GLDH activity of G4 and G5 were increased in vivo. Moreover, the ascorbic acid (AsA) and dehydroascorbate (DHA) contents were up-regulated in both leaves and tubers. However, the shoots of G1 were suppressed and its leaves were deformed. Additionally, the AsA contents in G1 leaves and tubers decreased by 28.8% and 10.3%, respectively. The GLDH activity in leaves treated with L-galactono-1,4-lactone (L-GalL) increased in all lines, as did the AsA and DHA contents. These results indicate that GLDH activity plays an important role in regulating the AsA level as well as the growth and development of potato plants.

CSN6, a subunit of the COP9 signalosome, is involved in early response to iron deficiency in Oryza sativa.

The COP9 signalosome (CSN) plays an important role in proteasome-mediated degradation by regulating CUL1 rubylation of the SCF ligase and is involved in many crucial biological processes. Here, we demonstrate a link between IDEF1 accumulation and the decline in COP9 derubylation activity in response to iron deficiency (-Fe) in rice (Oryza sativa). CSN6 expression is rapidly down-regulated during Fe depletion, contributing to reduced CSN activity, as judged by CSN5 and CUL1 expression, indicating CSN6 is involved in the early stage response of -Fe. In contrast to CSN6, the IDEF1 protein and expression of several iron uptake/utilisation-related genes are increased in response to -Fe. Thus, we constructed CSN6 transgenic sense and antisense lines and found that experimental depletion of CSN6 results in accumulation of the IDEF1 protein and up-regulation of several iron uptake/utilisation-related genes. Furthermore, IDEF1 can be decorated with K48-linked polyubiquitin and degraded via the 26S proteasome. Accumulated IDEF1 in antisense lines led to increased chlorophyll and Fe content in seedlings during -Fe. Collectively, the cellular CSN6 level is decreased during early stages of -Fe to ensure the rapid accumulation of IDEF1, which in turn up-regulates several iron uptake/utilisation-related genes to help overcome -Fe stress in rice.

Suppression of tomato SlGGP aggravates methyl viologen-mediated oxidative stress

Ascorbate (AsA) is an important antioxidant that can scavenge reactive oxygen species to protect plant cells against oxidative stress. Guanosine 5'-diphosphate (GDP)-L-galactose phosphorylase (GGP) is a key enzyme in the AsA biosynthetic pathway. To investigate the functions of GGP in AsA synthesis and oxidative stress tolerance in tomato, antisense lines with a reduced expression of SlGGP were obtained. Photobleaching after treatment of leaf disks with methyl viologen was more severe in transgenic lines compared to wild type (WT) plants. Moreover, compared with the WT plants, the transgenic plants showed a higher content of hydrogen peroxide, superoxide anion, malondialdehyde, as well as ion leakage, but a lower content of AsA and chlorophylls, ascorbate peroxidase activity, net photosynthetic rate, and maximal photochemical efficiency of photosystem II. Results of real-time quantitative polymerase chain reaction show that suppression of the SlGGP gene in the transgenic plants reduced their oxidative stress tolerance.

Regulation of Arabidopsis thaliana plasma membrane glucose-responsive regulator (AtPGR) expression by A. thaliana storekeeper-like transcription factor, AtSTKL, modulates glucose response in Arabidopsis

Biochemical, genetic, physiological, and molecular research in plants has demonstrated a central role of glucose (Glc) in the control of plant growth, metabolism, and development, and has revealed networks that integrate light, stresses, nutrients, and hormone signaling. Previous studies have reported that AtPGR protein as potential candidates for Glc signaling protein. In the present study, we characterized transcription factors that bind to the upstream region of the AtPGR gene isolated using the yeast one-hybrid screening with an Arabidopsis cDNA library. One of the selected genes (AtSTKL) appeared to confer elevated sensitivity to Glc response. Overexpression of AtSTKLs (AtSTKL1 and AtSTKL2) increased the sensitivity to Glc during the post-germination stages. In contrast, atstkl1 and atstkl2 antisense lines displayed reduced sensitivity to high Glc concentration during the early seedling stage. Furthermore, we showed that the two AtSTKLs bind to the 5′-GCCT-3′ element of the upstream promoter region of the AtPGR gene in vitro and repress the beta-glucuronidase (GUS) activity in AtPGR promoter-GUS (P999-GUS) transgenic plants. Green fluorescent protein (GFP)-tagged AtSTKLs were localized in the nuclei of transgenic Arabidopsis cells. Collectively, these results suggest that AtSTKL1 and AtSTKL2 function both as repressors of AtPGR transcription and as novel transcription factors in the Glc signaling pathway.

Proteomic analysis of mature barley grains from C-hordein antisense lines

Hordeins are the major storage proteins in barley grains and are responsible for their low nutritional quality. Previously, antisense C-hordein barley lines were generated and were shown to contain a more balanced amino acid composition and an altered storage protein profile. In the present study, a proteomic approach that combined two-dimensional gel electrophoresis (2-DE) and mass spectrometry was used to (1) identify the changes in the protein profile of non-storage proteins (salt soluble fraction) in antisense C-hordein barley lines (L1, L2 and L3) and (2) map the differentially expressed proteins compared to the non-transgenic control line (Hordeum vulgare cv. Golden Promise). Moreover, the changes in the proteins were correlated with the more balanced amino acid composition of these lines, with special attention to the lysine content. The results showed that suppression of C-hordein expression does not exclusively affect hordein synthesis and accumulation. The more balanced amino acid composition observed in the transgenic lines L1, L2 and L3 was an indirect result of the profound alterations in the patterns of the non-storage proteins. The observed changes included up-regulated expression of the proteins involved in stress and detoxification (L1), defence (L2 and L3), and storage globulins (L1, L2 and L3). To a lesser extent, the proteins involved in grain metabolism were also changed. Thus, the increased essential amino acids content results from changes in distinct protein sources among the three antisense C-hordein lines analyzed, although the up-regulated expression of lysine-rich proteins was consistently observed in all lines.

Control of potato tuber dormancy and sprouting by expression of sense and antisense genes of pyrophosphatase in potato

Inorganic pyrophosphate (PPi) is an enzyme involved in sugar metabolism in potato tubers. In our previous study, we isolated an inorganic pyrophosphatase (PPase) gene from potato and obtained the transgenic potato plants transformed with the sense and antisense PPase genes respectively. In the present experiment, the physiological indexes, tuber dormancy, and sprouting characteristics of the transgenic potatoes were analyzed and evaluated. The result showed that the PPase activity and the inorganic phosphate content of tubers were lower in the antisense transgenic plant lines but were higher in the sense transgenic plant lines, compared with wild-type tubers. Soluble sugars, such as glucose, fructose and sucrose increased in transgenic plants that had overexpression of the sense PPase gene, but decreased in the antisense transgenic plant lines, compared with wild-type tubers. Tuber sprouting time of the antisense transgenic plants were delayed for 2 and 3 weeks and reached the 100 % sprouting rate only after 14 and 16 weeks storage compared with the wild-type when tubers are stored under 25 and 4 °C, respectively. In contrast, tuber sprouting time of the sense transgenic plants was earlier by approximately 2 weeks than that of wild-type tubers under these storage temperatures.

The Essential Role of Cytokinin Signaling in Root Apical Meristem Formation during Somatic Embryogenesis.

The Essential Role of Cytokinin Signaling in Root Apical Meristem Formation during Somatic Embryogenesis.

Suppressing Sorbitol Synthesis Substantially Alters the Global Expression Profile of Stress Response Genes in Apple (Malus domestica) Leaves.

Sorbitol is a major product of photosynthesis in apple (Malus domestica) that is involved in carbohydrate metabolism and stress tolerance. However, little is known about how the global transcript levels in apple leaves respond to decreased sorbitol synthesis. In this study we used RNA sequencing (RNA-seq) profiling to characterize the transcriptome of leaves from transgenic lines of the apple cultivar 'Greensleeves' exhibiting suppressed expression of aldose-6-phosphate reductase (A6PR) to gain insights into sorbitol function and the consequences of decreased sorbitol synthesis on gene expression. We observed that, although the leaves of the low sorbitol transgenic lines accumulate higher levels of various primary metabolites, only very limited changes were found in the levels of transcripts associated with primary metabolism. We suggest that this is indicative of post-transcriptional and/or post-translational regulation of primary metabolite accumulation and central carbon metabolism. However, we identified significantly enriched gene ontology terms belonging to the 'stress related process' category in the antisense lines (P-value < 0.05). These include genes involved in the synthesis/degradation of abscisic acid, salicylic acid and jasmonic acid, nucleotide-binding site leucine-rich repeat (NBS-LRR) disease resistance genes and ATP-binding cassette (ABC) transporter genes. This suggests that sorbitol plays a role in the responses of apple trees to abiotic and biotic stresses.

Modifying glycoalkaloid content in transgenic potato – Metabolome impacts

Metabolite profiling has been used to assess the potential for unintended composition changes in potato (Solanum tuberosum L. cv. Desirée) tubers, which have been genetically modified (GM) to reduce glycoalkaloid content, via the independent down-regulation of three genes SGT1, SGT2 and SGT3 known to be involved in glycoalkaloid biosynthesis. Differences between the three groups of antisense lines and control lines were assessed using liquid chromatography–mass spectrometry (LC–MS) and gas chromatography (GC)–MS, and data analysed using principal component analysis and analysis of variance. Compared with the wild-type (WT) control, LC–MS revealed not only the expected changes in specific glycoalkaloid levels in the GM lines, but also significant changes in several other metabolites, some of which were explicable in terms of known pathways. Analysis of polar and non-polar metabolites by GC–MS revealed other significant (unintended) differences between SGT lines and the WT, but also between the WT control and other control lines used.

Effects of Ca&lt;sup&gt;2+&lt;/sup&gt; and polyethylene glycol on the chlorophyll fluorescence parameters of transgenic OsCaS rice (Oryza sativa L.)

Ca2+ is an important factor mediating many biotic and abiotic stress responses in plants. In this study, we measured the chlorophyll (Chl) fluorescence of transgenic rice with increased or decreased expression of a calcium-sensing receptor (OsCaS) gene during water deficit caused by polyethylene glycol to prove our hypothesis that increased Ca2+ in combination with increased OsCaS could enhance the drought resistance of transgenic rice. Transcript abundance (evaluated by RT-PCR) was significantly lower in OsCaS antisense line 766 (AS766) than that in the wild type, while the overexpression line 777 (O777) showed four times higher amount than that in the wild type. Chl fluorescence showed that the photochemical quantum yield of PSII in the light increased due to addition of Ca2+ in the O777, but dropped in the AS766. Nonphotochemical quenching increased under stress in both transgenic lines and in the wild type, but less in the O777. Nonregulatory quantum yield of energy dissipation showed no significant change under drought stress. Photochemical quenching was significantly higher in the O777 than those in the AS766 and in the wild type after the Ca2+ treatment. In the absence of stress, the electron transport rate (ETR) was significantly higher in the O777 than in both the AS766 and the wild type. In contrast, the ETR of the wild type and both transgenic lines decreased under drought stress, while the effect of polyethylene glycol was partially alleviated by Ca2+ addition in the O777. In summary, excitation energy conversion and dissipation by PSII were regulated by Ca2+ in the O777. It might partially alleviate the effect of drought stress, whereas addition of Ca2+ had no effect in the wild type and the AS766.

VDAC2 involvement in the stress response pathway in Arabidopsis thaliana

The voltage-dependent anion channel (VDAC) is the major transport protein in the outer membrane of mitochondria, and is involved in the formation of a permeable transition pore and metabolite transport. In this study, we explored the role of Arabidopsis thaliana VDAC2 (AtVDAC2) in the signal transduction pathway in Arabidopsis under salt stress. We investigated the germination rates of AtVDAC2 transgenic lines under salicylic acid (SA) treatment, and found that AtVDAC2 can affect the sensitivity of Arabidopsis to SA. Furthermore, the stomatal apertures of AtVDAC2 transgenic Arabidopsis were calculated. Results showed that the over-expression lines showed the obvious stomatal closure, while in the antisense lines, no obvious changes in stomatal apertures were found. In addition, we also detected the expression levels of salt stress and SA response-associated genes in transgenic plants and found that AtVDAC2 affected the expression of these genes. Our study investigated the role of AtVDAC2 in SA and salt stress response in Arabidopsis; our observations provide some helpful information for better understanding the direct and downstream functions of AtVDAC.

Impacts on the metabolome of down-regulating polyphenol oxidase in potato tubers.

Tubers of potato (Solanum tuberosum L. cv. Estima) genetically modified to reduce polyphenol oxidase (PPO) activity and enzymatic discolouration were assessed for changes in the metabolome using Liquid Chromatography-Mass Spectrometry (LC-MS) and Gas Chromatography (GC)-MS. Metabolome changes induced over a 48 hour (h) period by tuber wounding (sliced transverse sections) were also assessed using two PPO antisense lines (asPPO) and a wild-type (WT) control. Data were analysed using Principal Components Analysis and Analysis of Variance to assess differences between genotypes and temporal changes post-tuber wounding (by slicing). The levels of 15 metabolites (out of a total of 134 that were detected) differed between the WT and asPPO lines in mature tubers at harvest. A considerably higher number (63) of these metabolites changed significantly over a 48 h period following tuber wounding. For individual metabolites the magnitude of the differences between the WT and asPPO lines at harvest were small compared with the impacts of tuber wounding on metabolite levels. Some of the observed metabolite changes are explicable in terms of pathways known to be affected by wound responses. Whilst some statistically significant interactions (11 metabolites) were observed between line and time after wounding, very few profiles were consistent when comparing the WT with both asPPO lines, and the underlying metabolites appeared to be random in terms of the pathways they occupy. Overall, mechanical damage to tubers has a considerably greater impact on the metabolite profile than any potential unintended effects resulting from the down-regulation of PPO gene expression.

Over-expression of the MxIRT1 gene increases iron and zinc content in rice seeds.

Iron and zinc are essential in plant and human nutrition. Iron deficiency has been one of the causes of human mortality, especially in developing countries with high rice consumption. MxIRT1 is a ferrous transporter that has been screened from an iron-efficient genotype of the apple tree, Malus xiaojinensis Cheng et Jiang. In order to produce Fe-biofortified rice with MxIRT1 to solve the Fe-deficiency problem, plant expression vectors of pCAMBIA1302-MxIRT1:GFP and pCAMBIA1302-anti MxIRT1:GFP were constructed that led to successful production of transgenic rice. The transgenic plant phenotypes showed that the expression of endogenous OsIRT1 was suppressed by anti-MxIRT1 in antisense lines that acted as an opposing control, while sense lines had a higher tolerance under Zn- and Fe-deficient conditions. The iron and zinc concentration in T3 seeds increased by three times in sense lines when compared to the wild type. To understand the MxIRT1 cadmium uptake, the MxIRT1 cadmium absorption trait was compared with AtIRT1 and OsIRT1 in transgenic rice protoplasts, and it was found that MxIRT1 had the lowest Cd uptake capacity. MxIRT1 transgenic tobacco-cultured bright yellow-2 (BY-2) cells and rice lines were subjected to different Fe conditions and the results from the non-invasive micro-test technique showed that iron was actively transported compared to cadmium as long as iron was readily available in the environment. This suggests that MxIRT1 is a good candidate gene for plant Fe and Zn biofortification.

Function Analysis of &amp;lt;italic&amp;gt;PHO1&amp;lt;/italic&amp;gt; Antisense Transcript

Phosphorus is an essential macro element for plant growth and development. PHO1 is an important factor for phosphorus transportation from underground to aboveground parts. Different from Arabidopsis , rice PHO1 has a natural cis -antisense transcript. To clarify whether PHO1 antisense transcript plays a role in plant response to phosphorus stress, the antisense transcript of PHO1-2 was cloned and its response to P deficiency was analyzed. Transgenic plants with overexpression of the PHO1 antisense transcript were obtained. The results of phosphorus content measurement showed significantly lower phosphorus content in leaves of transgenic seedlings than that of the wild type, but the phosphorus content in the root of overexpression transgenic lines was higher than that of the wild type. This result was in consistent with the phenotype of PHO1 loss of function material. Quantitative RT-PCR results showed that the transcript level of PHO1 was significantly lowered in the antisense transcript overexpression transgenic lines. The results suggested that PHO1 antisense transcripts were involved in phosphorus homeostasis regulation by regulating the transcript level of PHO1 .

The transcription factor AREB1 regulates primary metabolic pathways in tomato fruits

Tomato fruit development is regulated both by the action of plant hormones and by tight genetic control. Recent studies suggest that abscisic acid (ABA) signalling may affect different aspects of fruit maturation. Previously, it was shown that SlAREB1, an ABA-regulated transcription factor involved in stress-induced responses, is expressed in seeds and in fruit tissues in tomato. Here, the role of SlAREB1 in regulating the expression of genes relevant for primary metabolic pathways and affecting the metabolic profile of the fruit was investigated using transgenic tomato lines. Metabolite profiling using gas chromatography-time of flight mass spectrometry (GC-TOF-MS) and non-targeted liquid chromatography-mass spectrometry (LC-MS) was performed on pericarp tissue from fruits harvested at three stages of fruit development. Principal component analysis of the data could distinguish the metabolite profiles of non-transgenic fruits from those that overexpress and down-regulate SlAREB1. Overexpression of SlAREB1 resulted in increased content of organic acids, hexoses, hexose-phosphates, and amino acids in immature green, mature green, and red ripe fruits, and these modifications correlated with the up-regulation of enzyme-encoding genes involved in primary carbohydrate and amino acid metabolism. A non-targeted LC-MS analysis indicated that the composition of secondary metabolites is also affected in transgenic lines. In addition, gene expression data revealed that some genes associated with fruit ripening are also up-regulated in SlAREB1-overexpressing lines compared with wild-type and antisense lines. Taken together, the results suggest that SlAREB1 participates in the regulation of the metabolic programming that takes place during fruit ripening and that may explain part of the role of ABA in fruit development in tomato.

Molecular and Cellular Characterization of the Tomato Pollen Profilin, LePro1

Profilin is an actin-binding protein involved in the dynamic turnover and restructuring of the actin cytoskeleton in all eukaryotic cells. We previously cloned a profilin gene, designated as LePro1 from tomato pollen. To understand its biological role, in the present study, we investigated the temporal and spatial expression of LePro1 during pollen development and found that the transcript was only detected at late stages during microsporogenesis and pollen maturation. Using antisense RNA, we successfully knocked down the expression of LePro1 in tomato plants using stable transformation, and obtained two antisense lines, A2 and A3 showing significant down-regulation of LePro1 in pollen resulting in poor pollen germination and abnormal pollen tube growth. A disorganized F-actin distribution was observed in the antisense pollen. Down-regulation of LePro1 also appeared to affect hydration of pollen deposited on the stigma and arrested pollen tube elongation in the style, thereby affecting fertilization. Our results suggest that LePro1 in conjunction with perhaps other cytoskeletal proteins, plays a regulatory role in the proper organization of F-actin in tomato pollen tubes through promoting actin assembly. Down-regulation of LePro1 leads to interruption of actin assembly and disorganization of the actin cytoskeleton thus arresting pollen tube growth. Based on the present and previous studies, it is likely that a single transcript of profilin gives rise to multiple forms displaying multifunctionality in tomato pollen.

Decrease of Auxin Binding Protein 1 Gene Expression Alters Shoot Development in Ramie

AUXIN BINDING PROTEIN1 (ABP1) has long been proposed as a candidate of auxin receptor to mediate auxin action in plants. It is essential for plant organ development. The function of ABP1 in the development of ramie (Boehmeria nivea L.) remains unclear. Here, we examined the role of the ABP1 gene in the development of shoot in ramie, especially in leaf venation ontogeny, by transforming ABP1 antisense construct into the plants. We showed that ABP1 expression was decreased in ABP1 antisense transgenic plantlets. The decrease of ABP1 expression led to defects in plant growth including dwarf plant, and decreased apical dominance in the intact plant. In addition, ABP1 antisense lines exhibited a serious retard of leaf development involving small and twisted leaves, sparse trichome, and a decreased cell expansion. Moreover, the decreased ABP1 expression repressed the development of leaf venation. ABP1 antisense lines developed an incomplete leaf venation, which became smaller and lacked in some leaf tissues. Though the cell arrangement was normal in midrib and lateral vein, cell size was decrease and some cell layers were missing in midrib. The abnormal phenotype of leaf venation was attributed to the decreases in cell number and size. Our data support that ABP1 is necessary for shoot growth in ramie. More importantly, it plays a key role for the development of ramie leaf venation by regulating both cell expansion and division.