IN THE NAME OF GOD

LEAF SENESCENCE
Department of plant biotechnology Guilan university
Presentation: F.masoomi-aladizgeh Mail: masoomi.farhad2030@yahoo.com

Senescence is the age-dependent deterioration process at the cellular, tissue, organ, or organismal level, leading to death or the end of the life span. During senescence, leaf cells undergo rather orderly changes in cell structure, metabolism, and gene expression. The earliest and most significant change in cell structure is the breakdown of the chloroplast, the organelle that contains up to 70% of the leaf protein. Increased catabolic activity is responsible for converting the cellular materials accumulated during the growth phase of leaf into exportable nutrients that are supplied to developing seeds or to other growing organs.

Arabidopsis thaliana is the best model for genetic studying

Arabidopsis thaliana is a favorite model for the molecular genetic study of leaf senescence. It has a short life cycle. Its leaves undergo readily distinguishable developmental stages. It has one of the smallest genomes in the plant kingdom: 115,409,949 base pairs of DNA distributed in 5 chromosomes (2n = 10). It can be easily grown in the lab in a relatively small space.

Development is rapid. It only takes 5 6 weeks from seed germination to the production of a new crop of seeds.

It is a prolific producer of seeds (up to 10,000 per plant) making genetics studies easier. It is normally self-pollinated so recessive mutations quickly become homozyguos and thus expressed. It almost has a 25,498 genes, but the scientist have not located all genes yet.

Leaf senescence associated cell death as a programmed cell death

Programmed cell death (PCD) is a self-destructing cellular process triggered by external or internal factors and mediated through an active genetic program. Cell death does not occur coherently but starts with local patches of early-dying cells and then propagates into the whole-leaf area.

A few examples of PCD in plants are observed:

I. It is in the pathogen-induced hypersensitive response (HR). II. It is in terms of the biological function, PCD in leaf senescence is mostly for remobilization of nutrients from the leaf to other organs including developing seeds. III. Its role in germination-related degeneration of aleuron.

Structural and Biochemical Changes in Leaf Senescence-Associated Cell Death

A notable feature of cellular structural change during leaf senescence is the order of disintegration of intracellular organelles.

The earliest structural changes occur in the chloroplast, In contrast, the nucleus and mitochondria that are essential for gene expression and energy production, respectively, remain intact until the last stages of senescence.
The overall cellular content of polysomes and ribosomes decreases fairly early, reflecting a decrease in protein synthesis. This occurs concomitantly with reduced synthesis of rRNAs and tRNAs and mRNAs.

The senescence markers:

chlorophyll content Photochemical efficiency senescence-associated enzyme activities change of protein levels membrane ion leakage Gene expression RNase or peroxidase activity

Leaf senescence is accompanied by decreased expression of genes related to photosynthesis (CAB2) and protein synthesis (RPS, RBC) and by increased expression of senescence-associated genes (SAGs).

Microarray analysis, RT-PCR, ESTs and RNA gel blot analysis can indicate contents of gene expression.

Factors involved in leaf senescence

There are two factors:

1) Internal factors (include various phytohormones and reproductive development as well as developmental age). 2) External factors (include stresses such as high or low temperature, drought, ozone, nutrient deficiency, pathogen infection, and shading.)

Internal factors Hormones Cytokinin Ethylene Auxin JA ABA SA GA Reproduction

Regulatory network Developmental age

External factors Abiotic UV-B or ozone Nutrient limitation Heat or cold Drought Biotic Shading Pathogen attack or wounding

1
Macromolecule degradation Nutrient salvage & translocation Detoxification & defense Chlorophyll loss/ nitrogen and lipid mobilization Increase of antioxidants and defense-related genes Cell death DNA laddering/ disruption of the nucleus and mitochondria Disintegration of the plasma and vacuolar membranes

Involvement of Phytohormone Pathways in Leaf Senescence

The hormonal pathways appear to play at all the stages of leaf senescence, including the initiation phase of senescence, progression, and the terminal phases.

Each plant hormone affects various developmental and/or environmental events in a complex manner. This causes difficulties in assaying the roles of the hormonal pathways in leaf senescence.

Cytokinin:
The endogenous cytokinin level drops during leaf senescence and exogenous application or endogenous enhancement of cytokinin content using the senescencespecific SAG12 promoter delays senescence.

The genes involved in cytokinin synthesis, a cytokinin synthase and adenosine phosphate isopentenyltransferase (IPT) genes, are downregulated and a gene for cytokinin degradation, cytokinin oxidase, is upregulated in senescing leaves.

Ethylene:
ethylene is an important positive regulator of leaf senescence. The ethylene biosynthetic genes encoding ACC synthase, ACC oxdiase, and nitrilase are upregulated in senescing leaves.

Arabidopsis mutants, ethylene-resistant 1 (etr1) and ethylene-insensitive 2 (ein2), that are deficient in ethylene perception and signal transduction, respectively, exhibited significant delays in leaf senescence.

In transgenic Arabidopsis and tomato plants that constitutively overproduce ethylene do not exhibit earlier-onset leaf senescence, suggesting that ethylene alone is not sufficient to initiate leaf senescence. The old1 mutant displays a phenotype with earlier-onset senescence in an age-dependent manner.

The early-senescence phenotype was further accelerated by exposure to ethylene, showing that the old1 mutation resulted in alternation of both of the age- and ethylene signaling dependent leaf senescence.

 However, in the old1etr1 double mutant where ethylene perception was blocked by the mutation in the ETR1 gene, age-dependent earlier-onset leaf senescence still occurred but was not further accelerated by ethylene treatment.

Abscisic acid(ABA):
The ABA level increases in senescing leaves and exogenously applied ABA induces expression of several SAGs.

In some stress conditions ABA content increases in leaves, because the genes encoding the key enzyme in ABA biosynthesis, 9-cisepoxycarotenoid dioxygenase (NECD), and two aldehyde oxidase genes AAO1 and AAO3 show increased expression.

A recent report argued that ABA induces accumulation of H2O2 in senescing rice leaf, which in turn accelerates leaf senescence.

ABA also induces expression of antioxidant genes and enhances the activities of antioxidative enzymes such as superoxide dismutase (SOD), ascorbate peroxidase (APOD), and catalase (CAT).

Methyl jasmonate (MeJA):

Methyl jasmonate (MeJA) and its precursor JA promote senescence in detached oat (Avena sativa) leaves.

Exogenously applied MeJA to detached Arabidopsis leaves leads to a rapid loss of chlorophyll content and photochemical efficiency of photosystem II (PSII) and increased expression of SAGs such as SEN4, SEN5, and VPE.

They observed that JA-dependent senescence is defective in the JA-insensitive mutant coronatine insensitive 1 (coi1), implying that the JA signaling pathway is required for JA to promote leaf senescence.

Salicylic asid(SA):
A recent intriguing discovery in leaf senescence was the role of SA in age-dependent leaf senescence.

The concentration of endogenous SA is four times higher in senescing leaves of Arabidopsis.

SA involved in upregulation of several SAGs during leaf senescence such as PR1a, chitinase, and SAG12

Leaves from Arabidopsis pad4 mutants that are defective in the SA signaling pathway do not appear to undergo cell death as efficiently as the wild type.

Gibberellin asid(GA):
it has been reported that the gibberellins can retard leaf senescence.

In 17 species studied, only Taraxacumn officinaile demonstrated a gibberellic acid regulation of senescence.
Therefore, GA is the hormone that negatively regulates the leaf senescence.

IAA:
The auxin level increases during leaf senescence. Consequently, IAA biosynthetic genes encoding tryptophan synthase (TSA1), IAAld oxidase (AO1), and nitrilases (NIT1-3) are upregulated during age-dependent leaf senescence. Exogenous application of auxin represses transcription of some SAGs.

This implies that the auxin level increases during leaf senescence due to increased expression of auxin biosynthetic genes, which leads to delayed leaf senescence.

AUXIN RESPONSE FACTOR 2 (ARF2) is one of the transcription repressors in the auxin signaling pathway. Microarray analysis shows that expression of the ARF2 gene is induced in senescing leaves.

Disruption of ARF2 by T-DNA insertion causes delay in leaf senescence. The phenotype canonically puts ARF2 as a positive regulator of leaf senescence. Besides the regulatory genes mentioned above, several other regulatory genes of leaf senescence have been identified such as: DLS1, ORE1, ORE7, ORE9, SOR12

Division of phytohormones:
Ethylene

Enhancers for SAG expression

ABA MeJA SA

Inhibitors for SAG expression

Cytokinin IAA GA

Conclusion:
With the aid of microarray, we now know that more than 800 genes are distinctively upregulated during senescence, which illustrates the dramatic alteration in cellular physiology that underlies leaf senescence. It is also unlikely that all the cells within an individual leaf undergo coherent cell death. To better understand the senescence process, it will be necessary to develop assays that can monitor senescence symptoms and senescenceassociated cell death symptoms at the individual cellular level.

Refrence:
Pyung Ok Lim, Hyo Jung Kim, and Hong Gil Nam
Department of Science Education, Cheju National University, Jeju, Jeju, 690-756, Korea
Division of Molecular Life Sciences and National Core Research Center for Systems Bio-Dynamics, POSTECH, Pohang, Kyungbuk, 790-784, Korea;

email: nam@postech.ac.kr