Rubber Tree (Hevea brasiliensis) Bark Necrosis Syndrome I: Still No Evidence of a Biotic Causal Agent.

The main constraint in rubber plantations worldwide is the cessation of latex production because of two syndromes: (i) tapping panel dryness (TPD), a reversible physiological response to overexploitation; and (ii) bark necrosis (BN), an irreversible syndrome spreading from the collar toward the tapping cut. Early BN symptoms develop in the inner phloem tissues but never affect the cambium. Necrotic patches appear in the outer phloem, inducing bark cracking and peeling, but these alterations never lead to tree death. BN spreads gradually to neighboring rubber trees, and evidence of linear disease centers suggest that a pathogen may be involved, possibly transmitted by the tapping knife. Several previous etiological investigations (fungi, phytoplasma, bacteria, and virus) were performed (3) on leaves, bark, and latex using different methods (e.g., isolation, transmission, chemical treatments, and optic and electron microscope studies). Recent works focused on mechanically transmissible pathogens, such as viroid (2) or virus/double strand RNA, using RNA extraction (nonionic cellulose and appropriate ethanol concentrations) and treatment with RNase A, followed with sequential polyacrylamide gel electrophoresis (s-PAGE), reverse transcription-polymerase chain reaction (RT-PCR), degenerate oligonucleotide primer-PCR (DOP-PCR), and cloning and sequence analysis. While numerous viroid-like (between 250 and 400 nucleotides) and double strand virus-like (1,800 bp) low-molecular-weight RNAs were observed, no definite correlation was found with the BN status of trees. Sequencing of the various isolated RNAs only identified plasmids, nonpathogenic bacteria and yeasts, but none of the suspected pathogens. In addition, previous and recent transmission trials (tapping knife disinfection, bud grafting, bark implantation, and etc.) failed to confirm the involvement of a biotic agent. In conclusion, since these etiological investigations were inconclusive, a physiological disease is now suspected that involves exogenous stresses, nonoptimal vascular relations at the rootstock/scion junction and impaired cyanide metabolism (1,4). References: (1) H. Chrestin et al. Plant Dis. 88:1047, 2004. (2) N. Duran-Vila et al. J. Gen. Virol. 69:3069, 1988. (3) D. Nandris et al. Eur. J. For. Pathol. 21:340, 1991. (4) D. Nandris et al. Plant Dis. 88:1047, 2004.

Rubber Tree (Hevea brasiliensis) Bark Necrosis Syndrome II: First Comprehensive Report on Causal Stresses.

Bark necrosis (BN), described and first studied in Côte d'Ivoire in the 1980s (2), affects most modern rubber plantations (i.e., grafted trees with high-yielding clones, intensive exploitation due to tapping frequency, and use of Ethrel as a yield stimulant) worldwide with a wide range of severity across sites. While previous (3) and recent (4) etiological analyses remain inconclusive, environmental factors were shown associated with BN. Numerous epidemiological surveys conducted in various African and Asian plantations on recently tapped blocks (less than 10-year-old trees) revealed the nonrandom location of the earliest single diseased trees. These risky areas are mainly characterized by the proximity of a swamp, plantation road, windrow, old bulldozer track, residual forest stump, or slope break. In BN emergence areas, while no significant correlation was found with chemical soil parameters, physical soil analyses (e.g., penetrometer) revealed higher soil compaction often associated with poorer rhizogenesis in BN trees (comparative root counts made in pits close to healthy or BN trees). Furthermore, initial BN symptoms were preferentially observed near the grafted bud at the rootstock/scion junction (RS/S). Numerous comparative ecophysiological measurements of leaf water potential, stem water potential, and predawn base potential using a plant moisture stress (PMS) pressure chamber indicated water stress in BN trees. These results and preliminary dye transfer studies at the RS/S junction suggested a nonoptimal vascular relation between the root system and the trunk of BN trees. In conclusion, compaction-associated reduced water availability of the soil and poor root capacity to meet the water demand during drier dry seasons combined with disturbed sap flows and recurrent local water drainage (latex flows) are now suspected to jointly act as the main exogenous causal stresses that induce the BN process at the RS/S bud zone before spreading upward to the tapping cut. This multidisciplinary approach gives a new comprehensive scenario for the emergence of this multifactorial physiological disease, now suspected to involve cyanide release (1) into the inner phloem of the rubber tree. References: (1) H. Chrestin. Plant Dis. 88:1047, 2004. (2) D. Nandris et al. Eur. J. For. Pathol. 21:325, 1991. (3) D. Nandris et al. Eur. J. For Pathol. 21:340, 1991. (4) F. Pellegrin et al. Plant Dis. 88:1046, 2004.

Rubber Tree (Hevea brasiliensis) Bark Necrosis Syndrome III: A Physiological Disease Linked to Impaired Cyanide Metabolism.

First attempts to discriminate between tapping panel dryness (TPD) and bark necrosis (BN), two Hevea sp. bark diseases leading to the cessation of latex production, showed differences in latex biochemical characteristics (1). Further, contrary to TPD, BN is characterized by inner phloem necrosis starting at the rootstock/scion junction (RS/S) and spreading upward to the tapping cut. Recent etiological (3) and epidemiological studies did not provide evidence of a causative pathogen for BN, but showed that BN is favored by a combination of various stresses (2). Searching for molecular markers of BN using sodium dodecyl sulfate-polyacrylamide gel electrophoresis analyses highlighted differential expression of some proteins in the latex and bark, especially a 67-kDa protein, which accumulates in the inner phloem of the BN trees. This protein was identified by peptide microsequencing as a linamarase (cyanogenic ß-glucosidase). This led to the suspicion of the involvement of cyanogenesis in the spread of the syndrome inside the inner bark. The cDNAs of enzymes involved in cyanide (CN) metabolism (linamarase, hydroxynitrile lyase, and cyanoalanine synthase) were cloned from our Hevea sp. phloem specific cDNA library. In addition, the most BN-susceptible rubber clones were shown to exhibit higher cyanide potentions in the leaves and bark, together with low cyanoalanine synthase (CAS) gene expression and activity. Furthermore, linamarine (the cyanogene glucoside substrate of linamarase) was shown to accumulate in the phloem at the base of the trunk, especially above the rootstock/scion junction. The results of biochemical and gene expression studies associated with recent ecophysiological advances (2) strongly suggest a possible cell decompartmentalization near the RS/S junction, resulting in a local release of toxic concentration of highly diffusive CN. This, combined with a lethal imbalance between cyanogenic and CN-detoxifying activities (CAS) in the phloem of BN trees, could lead to poisoning of neighboring cells and to the spread of tissue necrosis toward the tapping cut. In conclusion, after providing evidence of exogenous factors favoring BN (2), this report highlights endogenous disorders that may be at the origin of this physiological disease leading to BN. References: (1) D. Nandris et al. Eur. J. For. Pathol. 21:325, 1991. (2) D. Nandris et al. Plant Dis. 88:1047, 2004.