Functional Diversity of Class XI Myosins in Arabidopsis thaliana.

Plant myosin XI acts as a motive force for cytoplasmic streaming through interacting with actin filaments within the cell. Arabidopsis thaliana (At) has 13 genes belonging to the myosin XI family. Previous reverse genetic approaches suggest that At myosin XIs are partially redundant, but are functionally diverse for their specific tasks within the plant. However, the tissue-specific expression and enzymatic properties of myosin XIs have to date been poorly understood, primarily because of the difficulty in cloning and expressing large myosin XI genes and proteins. In this study, we cloned full-length cDNAs and promoter regions for all 13 At myosin XIs and identified tissue-specific expression (using promoter-reporter assays) and motile and enzymatic activities (using in vitro assays). In general, myosins belonging to the same class have similar velocities and ATPase activities. However, the velocities and ATPase activities of the 13 At myosin XIs are significantly different and are classified broadly into three groups based on velocity (high group, medium group and low group). Interestingly, the velocity groups appear roughly correlated with the tissue-specific expression patterns. Generally, ubiquitously expressed At myosin XIs belong to the medium-velocity group, pollen-specific At myosin XIs belong to the high-velocity group and only one At myosin XI (XI-I) is classified as belonging to the low-velocity group. In this study, we demonstrated the diversity of the 13 myosin XIs in Arabidopsis at the molecular and tissue levels. Our results indicate that myosin XIs in higher plants have distinct motile and enzymatic activities adapted for their specific roles.

Measurement of enzymatic and motile activities of Arabidopsis myosins by using Arabidopsis actins.

There are two classes of myosin, XI and VIII, in higher plants. Myosin XI moves actin filaments at high speed and its enzyme activity is also very high. In contrast, myosin VIII moves actin filaments very slowly with very low enzyme activity. Because most of these enzymatic and motile activities were measured using animal skeletal muscle α-actin, but not plant actin, they would not accurately reflect the actual activities in plant cells. We thus measured enzymatic and motile activities of the motor domains of two Arabidopsis myosin XI isoforms (MYA2, XI-B), and one Arabidopsis myosin VIII isoform (ATM1), by using three Arabidopsis actin isoforms (ACT1, ACT2, and ACT7). The measured activities were different from those measured by using muscle actin. Moreover, Arabidopsis myosins showed different enzymatic and motile activities when using different Arabidopsis actin isoforms. Our results suggest that plant actin should be used for measuring enzymatic and motile activities of plant myosins and that different actin isoforms in plant cells might function as different tracks along which affinities and velocities of each myosin isoform are modulated.

Evaluation of milk quality in delivering sterilized milk with soft tank transportation system.

A new transportation system is proposed recently to improve the defects of liquid transportation by tank trucks. This method is called "soft tank transportation system"; a driver installs a sac-like container (soft tank), which is made from a tarpaulin with high-pressure resistant-waterproof zippers, in a general cargo vehicle. To evaluate the quality of sterilized milk by using the soft tank transportation system, ground and marine transportation for a long distance which took about 36 h from the shipper's loading to the receiver's unloading in a high-temperature summer season (average outside temperature was 33.4 degrees C) were carried out. Although the difference of milk temperature before and after the delivery varied from -0.7 to +1.4 degrees C, there was no difference in milk quality (fat, nonfat solids, total dissolved solids, and pH) and no coliform bacteria were detected. It can be evaluated that sterilized milk was carried in keeping good conditions by soft tank transportation system.