Construction of linkage maps is crucial for genetic studies and marker-assisted breeding programs. Recent advances in next generation sequencing technologies allow for the generation of high-density linkage maps, especially in non-model species lacking extensive genomic resources. Here, we constructed a high-density integrated genetic linkage map of rubber tree (Hevea brasiliensis), the sole commercial producer of high-quality natural rubber. We applied a genotyping-by-sequencing (GBS) technique to simultaneously discover and genotype single nucleotide polymorphism (SNP) markers in two rubber tree populations. A total of 21,353 single nucleotide substitutions were identified, 55% of which represented transition events. GBS-based genetic maps of populations P and C comprised 1704 and 1719 markers and encompassed 2041 cM and 1874 cM, respectively. The average marker densities of these two maps were one SNP in 1.23–1.25 cM. A total of 1114 shared SNP markers were used to merge the two component maps. An integrated linkage map consisted of 2321 markers and spanned the cumulative length of 2052 cM. The composite map showed a substantial improvement in marker density, with one SNP marker in every 0.89 cM. To our knowledge, this is the most saturated genetic map in rubber tree to date. This integrated map allowed us to anchor 28,965 contigs, covering 135 Mb or 12% of the published rubber tree genome. We demonstrated that GBS is a robust and cost-effective approach for generating a common set of genome-wide SNP data suitable for constructing integrated linkage maps from multiple populations in a highly heterozygous agricultural species.

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The tapping panel dryness (TPD) syndrome of rubber is characterized by the reduction or ultimately total cessation of latex flow upon tapping, due to physiological disorders in the bark tissue. The protein pattern in the cytoplasm from healthy and TPD tree latex cells was compared by electrophoresis. Two polypeptides (P15 and P22) of 15 and 22 kDa, respectively, were found to accumulate in the cytosol of the TPD-affected trees, whereas a 29 kDa polypeptide (P29) appeared de novo. P15 and P22 were identified as REF (Hev b1) and SRPP (Hev b3), respectively, two proteins proposed to be involved in rubber biosynthesis. P29 appeared to be a new member of the patatinlike protein family. Specific molecular probes were designed for a detailed characterization of REF and SRPP gene expression and RFLP mapping. This allowed the demonstration that REF and SRPP display very similar expression profiles. They are highly over-expressed by the tappinginduced metabolic activation, although not by wounding per se, or ethylene or ABA. In addition to this similarity in gene expression, they were found to share one common locus in the genome. No significant difference in REF and SRPP gene expression was observed between healthy and TPD trees, indicating that their TPD-related accumulation in the cytosol was not transcriptionally regulated. Western blot analysis demonstrated that osmotic lysis of the sedimentable organelles (lutoids) in vitro caused the release of REF and SRPP from the rubber particle membrane into the cytosol. A mechanism of cellular delocalization as a consequence of the lutoids instability is proposed to explain REF and SRPP accumulation in the cytosol of TPD trees.

This research aimed to scale up the production of starch-based super water absorbent (SWA) and to validate the practical benefits of SWA for agricultural applications. SWA was successfully prepared in an up-scaling production by radiation-induced graft polymerization of acrylic acid onto cassava starch. Chemical characterization by FTIR and thermal characterization by TGA showed results that differentiated starting materials from the prepared SWA, thus confirming effective preparation of starch-based SWA via radiation-induced graft polymerization. SEM results visibly revealed a highly porous morphology of the synthesized SWA, substantiating its high swelling ability. Results from the field tests, performed for two seasons, revealed that the prepared SWA was able to increase the survival rate of young rubber trees planted in arid area by up to 40%, while simultaneously enhancing the growth characteristics of the young rubber trees.