Programmed cell death occurs asymmetrically during abscission in tomato

Abscission occurs specifically in the abscission zone (AZ) tissue as a natural stage of plant development. Previously, we observed delay of tomato (Solanum lycopersicum) leaf abscission when the LX ribonuclease (LX) was inhibited. The known association between LX expression and programmed cell death (PCD) suggested involvement of PCD in abscission. In this study, hallmarks of PCD were identified in the tomato leaf and flower AZs during the late stage of abscission. These included loss of cell viability, altered nuclear morphology, DNA fragmentation, elevated levels of reactive oxygen species and enzymatic activities, and expression of PCD-associated genes. Overexpression of antiapoptotic proteins resulted in retarded abscission, indicating PCD requirement. PCD, LX, and nuclease gene expression were visualized primarily in the AZ distal tissue, demonstrating an asymmetry between the two AZ sides. Asymmetric expression was observed for genes associated with cell wall hydrolysis, leading to AZ, or associated with ethylene biosynthesis, which induces abscission. These results suggest that different abscission-related processes occur asymmetrically between the AZ proximal and distal sides. Taken together, our findings identify PCD as a key mechanism that occurs asymmetrically during normal progression of abscission and suggest an important role for LX in this PCD process.     

Changes in distribution of cell wall polysaccharides in floral and fruit abscission zones during fruit development in tomato (Solanum lycopersicum)

After fruit development has been triggered by pollination, the abscission zone (AZ) in the pedicel strengthens its adhesion to keep the fruit attached. Unpollinated flowers are shed at their respective AZs, whereas an enlargement of the same tissue is observed in pollinated flowers. After the fruit has developed and is fully ripened, shedding occurs easily at the AZ, indicating an acceleration of abscission. Cell wall degradation and synthesis may play important roles in these processes; however, little is understood. In this report, we have visualized changes in polysaccharide distribution in the AZs of pollinated versus unpollinated flowers and in the ripened fruits using immunohistochemistry. During floral abscission, a large increase was observed in LM15 labeling of xyloglucan specifically at the AZ in the abscising pedicel. LM5 and LM6 labeling of galactan and arabinan, respectively, also increased—LM5 throughout the pedicel and LM6 at the basal side of the AZ. The results suggest that xyloglucan, pectic galactan and arabinan play key roles in the abscission process. During fruit abscission, unlike in floral abscission, no AZ-specific cell wall polysaccharide deposition was observed; however, high autofluorescence was seen in the AZ of over-ripe fruit pedicels, suggesting secondary cell wall synthesis and lignification of the AZ prior to fruit abscission.     

Complexity and expression of the glutamine synthetase multigene family in the amphidiploid crop Brassica napus

In the amphidiploid genome of oilseed rape (Brassica napus) the diploid ancestral genomes of B. campestris and B. oleracea have been merged. As a result of this crossing event, all gene loci, gene families, or multigene families of the A and C genome types encoding a certain protein are now combined in one plant genome.In the case of the multigene family for glutamine synthetase, the key enzyme of nitrogen assimilation, six different cDNA sequences were isolated from leaf and root specific libraries. One sequence pair (BnGSL1/BnGSL2) was characterized by the presence of amino- terminal transit peptides, a typical feature of all nuclear encoded chloroplast proteins. Two other cDNA pairs (BnGSR1-1/BnGSR1-2 and BnGSR2-1/BnGSR2-2) with very high homology between each other were found in a root specific cDNA library and represent protein subunits for cytosolic glutamine synthetase isoforms.Comparative PCR amplifications of genomic DNA isolated from B. napus, B. campestris and B. oleracea followed by sequence–specific restriction analyses of the PCR products permitted the assignment of the cDNA sequences to either the A genome type (BnGSL1/BnGSR1- 1/BnGSR2-1) or the C genome type (BnGSL2/BnGSR1-2/BnGSR2-2). Consequently, the ancestral GS genes of B. campestris and B. oleracea are expressed simultaneously in oilseed rape. This result was also confirmed by RFLP (restriction fragment length polymorphism) analysis of RT-PCR products.In addition, the different GS genes showed tissue specific expression patterns which are correlated with the state of development of the plant material. Especially for the GS genes encoding the cytosolic GS isoform BnGSR2, a marked increase of expression could be observed after the onset of leaf senescence.     

Chilling-induced leaf abscission of Ixora coccinea plants. I. Induction by oxidative stress via increased sensitivity to ethylene

Exposing ixora ( Ixora coccinea ) plants to chilling temperatures (3–9°C for 3 days) resulted in increased leaf abscission, initiated 3 days after transfer to 20°C. Exposure to chilling also induced a 7-fold increase in ethylene production rates of abscission zone (AZ) tissue during the initial 5 h after chilling. The ethylene burst resulted from the high levels of 1-aminocyclopropane-1-carboxylic acid (ACC) accumulated in the AZ during the chilling period. ACC levels following chilling decreased also due to enhanced conjugation to 1-(malonylamino)cyclopropane-1-carboxylic acid (MACC). Treating plants prior to chilling with antioxidants, such as butylated hydroxyanisole (BHA), n -propyl gallate (PG), and vitamin E, significantly reduced chilling-induced leaf abscission. This effect was obtained despite the fact that ethylene production in the treated plants resembled that of chilled plants receiving no BHA. In addition, exposure of plants to ethylene (0.5–10 μl l⁻¹) for 1–3 days significantly enhanced leaf abscission only when they had been pre-chilled. These data imply that chilling-induced leaf abscission was closely correlated with increased sensitivity of the AZ to ethylene rather than with the chilling-induced ethylene burst. Based on the findings that the ethylene action inhibitor, 1-methylcyclopropene (1-MCP), and the antioxidant BHA inhibited both the chilling-induced and the ethylene-enhanced leaf abscission, it is concluded that: (1) although ethylene is essential for chilling-induced abscission, it is not the triggering factor; (2) oxidative processes derived from the chilling stress seem to be the trigger of chilling-induced leaf abscission, operating via increased sensitivity to ethylene.     

Involvement of hydrogen peroxide in leaf abscission signaling, revealed by analysis with an in vitro abscission system in Capsicum plants

Although auxin and ethylene play pivotal roles in leaf abscission, the subsequent signaling molecules are poorly understood. This is mainly because it is difficult to effectively treat the intact abscission zone (AZ) with pharmacological reagents. We developed an in vitro experimental system that reproduces stress-induced leaf abscission in planta. In this system, 1-mm-thick petiole strips, encompassing the AZ, were separated within 4 days of abscission at the AZ through cell wall degradation in an auxin depletion- and ethylene-dependent manner. The system allowed us to show that hydrogen peroxide (H(2)O(2)) is involved in abscission signaling. Microscopic analyses revealed continuous H(2)O(2) production by AZ cells. H(2)O(2) scavengers and diphenylene iodonium, an inhibitor of NADPH oxidase, suppressed in vitro abscission and cellulase expression. Conversely, the application of H(2)O(2) promoted in vitro abscission and expression of cellulase. Ethephon-induced abscission was suppressed by inhibitors of H(2)O(2) production, whereas the expression of ethylene-responsive genes was unaffected by both H(2)O(2) and an H(2)O(2) inhibitor. These results indicated that H(2)O(2) acts downstream from ethylene in in vitro abscission signaling. In planta, salinity stress induced the expression of genes that respond to ethylene and reactive oxygen species, and also induced H(2)O(2) production at the AZ, which preceded leaf abscission. These results indicate that H(2)O(2) has roles in leaf abscission associated with ethylene both in vitro and in planta.     

Three different polygalacturonases are expressed in tomato leaf and flower abscission, each with a different temporal expression pattern.

Abscission, or organ separation, is accompanied by a marked increase in hydrolases, which are responsible for the degradation of the middle lamella and the loosening of the primary cell wall surrounding cells in the separation layer. We recently reported on the cloning of a tomato (Lycopersicon esculentum) polygalacturonase (PG) cDNA, TAPG1, expressed during leaf and flower abscission. In addition to TAPG1, we have cloned two more PG cDNAs (TAPG2 and TAPG4) that are also expressed during leaf and flower abscission. The peptide sequences for the three abscission PGs are relatively similar (76-93% identity) yet different from the those of tomato fruit PG (38-41% identity). None of the three abscission PG mRNAs are expressed in fruit, stems, petioles, or anthers of fully open flowers. An RNase protection assay revealed that all three PGs are expressed in leaf and flower abscission zones and in pistils of fully open flowers. TAPG4 mRNA is detected much earlier than TAPG1 and TAPG2 mRNA during both leaf and flower abscission.     

两个大豆类受体蛋白激酶基因的克隆及其结构和功能的初步分析

利用高等植物类受体蛋白激酶基因的保守域设计简并引物的通过RT-PCR方法,从大豆叶片中克隆到两个新的,可能的类受体蛋白激酶基因的部分cDNA片段。对其基因结构的分析表明：在RLPK2的激酶保守域Vib与Ⅸ之间有一个407bp长的内含子。利用RT-PCR方法对它们的表达特性进行初步研究。发现这两个基因可能参与了对大豆叶片衰老和/或细胞分裂素延缓衰老过程的调节机制。