PhD

The project entitled “Biochemical and molecular studies on post-harvest physiological deterioration of cassava (Manihot esculenta Crantz)” was conducted at the Dept of Plant Physiology, College of Agriculture, Vellayani and at ICAR-CTCRI, Thiruvananthapuram during 2013 to 2015. The main objective of the work was to analyse the physiological biochemical and molecular mechanisms associated with post harvest physiological deterioration (PPD) and develop methods to delay the PPD in cassava.
The initial screening of the cassava genotypes for their performance of PPD was done with 61 different genotypes including some released varieties. Significant differences were observed for PPD development and shelf-life of harvested roots. Cassava genotypes such as IMS2-8, 9S-172, 11S-53, IRS 2-10 and 9S-286 started showing the visible discolouration in the parenchyma tissue earlier (less than 3 days of storage). Genotypes such as 9S-7, 9S-98, 11S-31, 11S-86, 11S-14, CE63-3, CI43-2, CR43-2, CR54-A5, CR59-8R, Sree Athulya and Kalpaka showed low PPD scores and better shelf-life. Tissue imprinting for peroxidase enzyme showed that there was a remarkable increase in peroxidase activity in the root tissues with increasing PPD symptoms. There was no correlation between the root morphological traits and PPD severity. Carotene and starch content of root did not influence the PPD in the genotypes studied. Chemotypic profile of roots with PPD symptoms was used to classify the genotypes based on PPD.
To develop an objective screening tool, Near Infrared Spectroscopy (NIRS) was utilized to analyse root samples for PPD. Principal component analysis (PCA) and chemometric tools clearly grouped the different PPD category in root tissues. Various stains such as saffranin, aniline blue, erythrocin, fast green and phloroglucinol stained the tissue specifically at vascular tissues and other cell components and were not suitable for detecting PPD.
Storage techniques such as storing the harvested roots in de-aerated bags, wax coating and burying the roots under the soil were employed with selected cassava varieties like Sree Athulya, Sree Jaya, Vellayani Hrashwa, Kalpaka and Sree Padmanabha to delay PPD. Wax coating was suitable to reduce PPD for few weeks. Effect of different storage temperature on PPD was studied for five cassava varieties. Root respiratory flux was higher in roots stored at ambient conditions compared to high (40o C) or low temperature (4o C) storage. There were significant positive correlation between root respiratory flux at 3 and 9 days of storage to the CAT and POX activities studied in different cassava varieties. Roots were treated with various food preservatives at two different concentrations (0.5 and 1%). There was a weak, but significant reduction in symptom development in butylated hydroxy touline – (BHT, at 0.5 and 1% level) treated roots compared to other treatments. Nearly three folds increase in total phenol content was noticed in BHT and butylated hydroxy anisole (BHA) treated roots. The plant hormones related to wound response such as Salicylic acid and jasmonic acid were used to study the PPD response in roots under storage. The roots did not show marked influence to hormone application.
Significant genetic variation was observed for PPD. The low PPD type genotypes such as 9S-7, 9S-98, 11S-31, 11S-86, 11S-14, CE63-3, CI43-2, CR43-2, CR54-A5, Sree Athulya and Kalpaka can be utilized for breeding programmes. High temperature storage of cassava at 40 oC resulted in reduced respiratory rate and increased antioxidant scavenging enzyme activity and also reduced the PPD. Differentiation of cassava roots at the metabolites level corresponding to visual symptoms and chemotypic profile of PPD and NIR spectroscopy offer a rapid screening tools. Among the different storage treatments, wax coating with antiseptic pre-treatment is most suitable and economical for increasing shelf-life of roots. Food preservatives like BHT and BHA have a significant, albeit marginal influence on PPD symptom development in cassava.