Genome-wide analysis of the CAD gene family reveals two bona fide CAD genes in oil palm.

Cinnamyl alcohol dehydrogenase (CAD) is the key enzyme for lignin biosynthesis in plants. In this study, genome-wide analysis was performed to identify CAD genes in oil palm (Elaeis guineensis). Phylogenetic analysis was then conducted to select the bona fide EgCADs. The bona fide EgCAD genes and their respective 5' flanking regions were cloned and analysed. Their expression profiles were evaluated in various organs using RT-PCR. Seven EgCAD genes (EgCAD1-7) were identified and divided into four phylogenetic groups. EgCAD1 and EgCAD2 display high sequence similarities with other bona fide CADs and possess all the signature motifs of the bona fide CAD. They also display similar 3D protein structures. Gene expression analysis showed that EgCAD1 was expressed most abundantly in the root tissues, while EgCAD2 was expressed constitutively in all the tissues studied. EgCAD1 possesses only one transcription start site, while EgCAD2 has five. Interestingly, a TC microsatellite was found in the 5' flanking region of EgCAD2. The 5' flanking regions of EgCAD1 and EgCAD2 contain lignin-associated regulatory elements i.e. AC-elements, and other defence-related motifs, including W-box, GT-1 motif and CGTCA-motif. Altogether, these results imply that EgCAD1 and EgCAD2 are bona fide CAD involved in lignin biosynthesis during the normal development of oil palm and in response to stresses. Our findings shed some light on the roles of the bona fide CAD genes in oil palm and pave the way for manipulating lignin content in oil palm through a genetic approach. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03208-0.

Application of arbuscular mycorrhizal fungi with Pseudomonas aeruginosa UPMP3 reduces the development of Ganoderma basal stem rot disease in oil palm seedlings.

The effect of arbuscular mycorrhizal fungi (AMF) in combination with endophytic bacteria (EB) in reducing development of basal stem rot (BSR) disease in oil palm (Elaeis guineensis) was investigated. BSR caused by Ganoderma boninense leads to devastating economic loss and the oil palm industry is struggling to control the disease. The application of two AMF with two EB as biocontrol agents was assessed in the nursery and subsequently, repeated in the field using bait seedlings. Seedlings pre-inoculated with a combination of Glomus intraradices UT126, Glomus clarum BR152B and Pseudomonas aeruginosa UPMP3 significantly reduced disease development measured as the area under disease progression curve (AUDPC) and the epidemic rate (R L) of disease in the nursery. A 20-month field trial using similar treatments evaluated disease development in bait seedlings based on the rotting area/advancement assessed in cross-sections of the seedling base. Data show that application of Glomus intraradices UT126 singly reduced disease development of BSR, but that combination of the two AMF with P. aeruginosa UPMP3 significantly improved biocontrol efficacy in both nursery and fields reducing BSR disease to 57 and 80%, respectively. The successful use of bait seedlings in the natural environment to study BSR development represents a promising alternative to nursery trial testing in the field with shorter temporal assessment.

Mating Compatibility and Restriction Analysis of Ganoderma Isolates from Oil Palm and Other Palm Hosts.

Mating compatibility and restriction analyses of Internal Transcribed Spacer (ITS) regions were performed to determine the relations between Ganoderma boninense, the most common species associated with basal stem rot in oil palm and Ganoderma isolates from infected oil palm, two ornamental palms, sealing wax palm (Cyrtostachys renda) and MacArthur palm (Ptychosperma macarthurii), an isolate from coconut stump (Cocos nucifera), Ganoderma miniatocinctum, Ganoderma zonatum and Ganoderma tornatum. The results showed that G. boninense was compatible with Ganoderma isolates from oil palm, G. miniatocinctum and G. zonatum, Ganoderma isolates from sealing wax palm, MacArthur palm and coconut stump. G. boninense was not compatible with G. tornatum. Therefore, the results suggested that the G. boninense, G. miniatocinctum, G. zonatum, and Ganoderma isolates from oil palm, ornamental palms and coconut stump could represent the same biological species. In performing a restriction analysis of the ITS regions, variations were observed in which five haplotypes were generated from the restriction patterns. An unweighted pair-group method with arithmetic averages (UPGMA) cluster analysis showed that all the Ganoderma isolates were grouped into five primary groups, and the similarity values of the isolates ranged from 97% to 100%. Thus, a restriction analysis of the ITS regions showed that G. boninense and the Ganoderma isolates from other palm hosts were closely related. On the basis of the mating compatibility test and the restriction analysis of the ITS regions performed in this study, a diverse group of Ganoderma species from oil palm and other palm hosts are closely related, except for G. tornatum and Ganoderma isolates from tea and rubber.

Symbiotic interaction of endophytic bacteria with arbuscular mycorrhizal fungi and its antagonistic effect on Ganoderma boninense.

Endophytic bacteria (Pseudomonas aeruginosa UPMP3 and Burkholderia cepacia UMPB3), isolated from within roots of oil palm (Elaeis guineensis Jacq.) were tested for their presymbiotic effects on two arbuscular mcorrhizal fungi, Glomus intraradices UT126 and Glomus clarum BR152B). These endophytic bacteria were also tested for antagonistic effects on Ganoderma boninense PER 71, a white wood rot fungal pathogen that causes a serious disease in oil palm. Spore germination and hyphal length of each arbuscular mycorrhizal fungal (AMF) pairing with endophytic bacteria was found to be significantly higher than spores plated in the absence of bacteria. Scanning electron microscopy (SEM) showed that the endophytic bacteria were scattered, resting or embedded on the surface hyaline layer or on the degraded walls of AMF spores, possibly feeding on the outer hyaline spore wall. The antagonistic effect of the endophytic bacteria was expressed as severe morphological abnormalities in the hyphal structures of G. boninense PER 71. The effects of the endophytic bacteria on G. boninense PER 71 hyphal structures were observed clearly under SEM. Severe inter-twisting, distortion, lysis and shriveling of the hyphal structures were observed. This study found that the effect of endophytic bacteria on G. intraradices UT126 and G. clarum BR152B resembled that of a mycorrhiza helper bacteria (MHB) association because the association significantly promoted AMF spore germination and hyphal length. However, the endophytic bacteria were extremely damaging to G. boninense PER 71.