Lateral root is primary organ for plant to explore and utilize soil nutrient efficiently. The development of lateral roots (LR) is controlled by both genetic factors and nutrient status in environment. To investigate the effects of nitrate (NO3-) on rice lateral root growth and nitrogen (N) uptake efficiency under upland condition, three treatments, including root-split culture and whole plant culture in N sufficient and deficient conditions, were used in a vermiculite culture experiment. Root-split treatment showed that the growth of lateral roots was stimulated by localized nitrate supply. However, in whole plant culture, elongation of lateral roots was induced by NO3- deficiency. The effects of NO3- on rice lateral root growth were genotype-dependent. Similar N concentration, soluble sugar concentration and N content in shoot were observed in both root-split treatment and whole plant culture under NO3- sufficient condition, suggesting that the nitrogen requirement for rice normal growth could be satisfied with only half of roots supplied with NO3-. In the root-split treatment, N uptake was positively correlated with the average of lateral root length (ALRL) in NO3--supplied side, suggesting that the ALRL is important for rice root N uptake in the environment where the nitrogen nutrient is limiting factor. No significant correlation was observed between N uptake and ALRL in whole plant culture under N sufficient condition, which implies that the length of lateral roots may not be the main factor to determine tire rice root N uptake in nutrient-rich zone. Morphological and metabolic evidence in this study provided some prospects for genetic improvement of root system characters to improve the efficiency of nutrient absorption in rice.
To understand the regulation system of nitrogen X-starvation in higher plants, a cDNA library from N-starved rice (Oryza sativa L.) seedlings was constructed using rapid subtraction hybridization (RaSH) procedure. Through reverse Northern analysis and Northern blotting, 18 unique known genes and two unique unknown genes were identified, which were up-regulated by N-starvation in rice. The known genes are involved in several metabolisms including carbon metabolism, secondary metabolite synthesis, ubiquitylation and protein degradation, phytohormone metabolism, signal transduction, growth regulator and transcription factors. Different induced expression patterns based on spatial and temporal express ions were found for these genes. The results indicate the cross-talks between N-starvation response and various metabolisms in plants.
Phosphorus is one of the three essential macroelements for plant growth. Plants respond to phosphorus starvation through adaptive mechanisms involved in morphological, biochemical and molecular changes. To investigate the molecular background of the adaptive mechanisms, the suppression subtractive hybridization (SSH) method was used to construct a rice phosphorus-starvation ( Pi-starvation) induced cDNA library. Through screening of the cDNA library and sequencing of the enriched cDNAs, 18 known genes and 47 novel genes were identified. The known genes are involved in different metabolic processes, including phosphate uptake and transport, signal transduction, protein synthesis and degradation, carbon metabolism and stress response. Northern analysis was performed to detect the expression patterns of some known genes and novel genes under different phosphorus levels. Different expression patterns of the selected genes were identified, which suggests that genes involved in different pathways may have different responses to Pi-starvation.
A vacuolar ATPase (V-ATPase.) B subunit gene has been cloned and characterized front a phosphorus starvation induced rice root subtractive cDNA library by suppression subtractive hybridization (SSH) method and RT-PCR amplification. This gene encodes a polypeptide of 487 amino acid residues, containing a conservative ATP binding site and with a molecular weight of 54.06 kD and an isoelectric point of 4.99, southern analysis of the. genomic DNA indicates that V-ATPase B subunit is encoded by a single gene in rice genome. The amino acid homologies of V-ATPase B subunits among different organisms range from 76% to 97% and reveals that the evolution of V-ATPase B subunit is accompanied with the biological evolution. Expression pattern analysis indicated that the maximal expression of V-ATPase B subunit gene occurred at an early stage (6 - 12 h) after phosphorus starvation in roots, and lately stage (24 - 48 It) in leaves. Under phosphorus deficiency, the up-regulated expression of V-ATPase gene was presumed to strengthen the proton transport and provide the required energy to maintain an electrochemical gradient across the tonoplast to facilitate Phosphorus transport.
