Analysis of phloem exudate collected from fruit-bearing stems of coconut palm: Palm trees as a source of molecules circulating in sieve tubes

Sieve tubes have been attracting widespread research interest because of their possible role in mediating physiological signals within the whole plant. However, progress in research into the function of sieve tubes has been limited by the low volume of sap available. To overcome this problem, we attempted to collect phloem exudate from tropical coconut palm trees (Cocos nucifera L. cv. Namhom). As much as 3 to 15 mL of exudate per hour was collected from the cut surface of the plant's fruit-bearing stem. Our analyses revealed that the characterized profiles of sugars (sucrose: 339 mM), amino acids (total concentration: 17.1 mM), cations (potassium: 48.3 mM), and proteins (total concentration: 0.1 /-lg /-lL-1) in the exudate were mostly consistent with those of phloem sap or phloem exudate collected from rice plants, castor bean plants, etc. This exudate was assumed to reflect the composition of the phloem sap from the source organs of coconut palm trees. The large volume of exudate collected contributed significantly to the analyses of the various compounds in the stream of sieve tubes.     

Immunohistochemical localization of metallothionein in plant tissues

The localization of metallothionein ( MT ) in the seeds and roots of soybean was investigated by immunohistochemistry. The germinating seeds at 2 hr, 1, 2, 3, 4 and 6 d including 1-mo root tips of soybean ( c.v. Toyosuzu ) with and without heavy metals ( Cu 400 μg Lor Zn 3 μg ml^?1) treatment were used to demonstrate the localization of MT by the indirect immunoperoxidase technique using polyclonal rabbit antirat MT conjugated to ascaris as a primary antibody. Metallothionein was localized in the proliferating regions such as the embryo in seeds, and root and shoot apices of both the control and heavy metals-treated plants. The intensity of MT staining in the proliferating regions generally increased as the soybean seeds germinate. Starting at about 1 day after germination, MT was found in the veins and vascular bundles suggesting its translocation to other organs. Similar observation hold true in the case of plants treated with heavy metals. This means that heavy metals treatment had no effect on MT localization. However, the heavy metals-treated plants showed higher concentration of MT over the control with respect to the growth stage of soybean seeds. These indicate that MT found in soybean plays a physiological role in heavy metal transport, detoxification and cell division in a similar manner to mammalian MT.