OsPIN1a Gene Participates in Regulating Negative Phototropism of Rice Roots

The complete open reading frame of OsPINla was amplified through reverse transcriptase- polymerase chain reaction （RT-PCR） based on the sequence deposited in GenBank to explore the relationship between the auxin efflux protein OsPINla and the negative phototropism of rice roots. Sequencing results showed that the GC content of OsPINla was 65.49%. The fusion expression vector pCAMBIA-1301-OsP/N1a：：GFP containing the OsPINla gene and a coding green fluorescent protein （gfp） gene was constructed. The fusion vector was transferred into onion epidermal cells by Agrobacterium tumefaciens transformation. The transient expression of OsPINla-GFP was mainly located in the nucleus and cell membrane. Moreover, the transgenic plants were obtained by Agrobacterium-mediated genetic transformation. Molecular detection performed by using PCR and β-glucuronidase staining showed that the target construct was integrated into the genome of rice. The negative phototropic curvatures of the transgenic rice roots were higher than those of the wild type. Similarly, the expression levels of OsPINla in the transgenic plants were considerably higher than those in the wild-type plants. These results suggest that OsPINla is crucial in the negative phototropic curvature of rice roots.

Effect of Indoleacetic acid (IAA) on the Negative Phototropism of Rice Root

To explore the effects of IAA on negative phototropism of rice ( Oryza sativa L.) root, agar block containing IAA was unilaterally applied on root tip to examine the phototropic response of root to exogenous IAA, and microstructure of the bending part was observed with an optical microscope. The growth of seminal roots could be regulated by exogenous IAA as well as light, as a result the root bent towards the site treated, causing asymmetric growth of the root cells at the elongation zone and consequently bending growth. IAA concentration in the shaded side of adventitious root increased much greater at 1.5 h after the start of irradiation. The unequal lateral IAA distribution can be concluded to be the main cause for negative phototropism of rice root.

Calcium Signaling is Involved in Negative Phototropism of Rice Seminal Roots

Calcium ions （Ca2＋） act as an intracellular second messenger and affect nearly all aspects of cellular life. They are functioned by interacting with polar auxin transport, and the negative phototropism of plant roots is caused by the transport of auxin from the irradiated side to the shaded side of the roots. To clarify the role of calcium signaling in the modulation of rice root negative phototropism, as well as the relationship between polar auxin transport and calcium signaling, calcium signaling reagents were used to treat rice seminal roots which were cultivated in hydroculture and unilaterally illuminated at an intensity of 100-200 pmol/（m2.s） for 24 h. Negative phototropism curvature and growth rate of rice roots were both promoted by exogenous CaCI2 lower than 100 pmol/L, but inhibited by calcium channel blockers （verapamil and LaCI3）, calcineurin inhibitor （chlorpromazine, CPZ）, and polar auxin transport inhibitor （N-l-naphthylphthalamic acid, NPA）. Roots stopped growing and negative phototropism disappeared when the concentrations increased to 100 pmol/L verapamil, 12.500 ~Jmol/L LaCI3, 60 pmol/L CPZ, and 6 pmol/L NPA. Moreover, 100 pmol/L CaCI2 could relieve the inhibition of LaCI3, verapamil and NPA. The enhanced negative phototropism curvature was caused by the transportation of more auxin from the irradiated side to the shaded side in the presence of exogenous Ca2＋. Calcium signaling plays a key role as a second messenger in the process of light signal regulation of rice root growth and negative phototropism.

Negative Phototropism of Chlorophytum comosum Roots and Their Mechanisms

The aerial roots of Chlorophytum comosum were grown hydroponically,allowing us to study the performance and mechanism of negative phototropism. The results of this study were as follows. All the adventitious roots and their branch roots bent away from light with a maximum curvature of approximately 88.5°. Blue-violet light prominently induced negative phototropism while red light had no effect. The root cap was the site of photo perception. Roots with shaded or divested root caps exposed to unilateral light showed no negative phototropism,but resumed their original characteristics when the shade was removed or when new root caps grew. The curvature increased when the light intensity ranged 0–110 μmol·m^(-2)·s^(-1). The negative phototropism curvature could be promoted by exogenous CaCl_2 but was inhibited by exogenous LaCl_3; exogenous CaCl_2 could reduce the inhibitory effect of LaCl_3. Unilateral light induced the horizontal transport of IAA from the irradiated side to the shaded side,resulting in an unequal distribution of IAA in both the sides,leading to negative phototropism. The horizontal transport of IAA was promoted by exogenous Ca^(2+) but inhibited by exogenous La^(3+).