OsRPK1, a novel leucine-rich repeat receptor-like kinase,negatively regulates polar auxin transport and root development in rice

Background

Leucine-rich-repeat receptor-like kinases (LRR-RLKs) represent the largest subfamily of putative RLKs in plants. Although several members in this subfamily have been identified, the studies about the relationships between LRR-RLKs and root development are still few. We previously identified a novel LRR-RLK in rice roots, and named it OsRPK1.

Methods

In this study, we first detected OsRPK1 kinase activity in vitro, and assessed its expression profile. We then investigated its biological function using transgenic rice plants over- and under-expressing OsRPK1.

Results

The OsRPK1 gene, which encodes a Ca^2 +-independent Ser/Thr kinase, was predominantly expressed in root tips, leaf blades, and undifferentiated suspension cells, and was markedly induced by treatment with auxin or ABA. Knockdown of OsRPK1 promoted the growth of transgenic rice plants, and increased plant height and tiller numbers. In contrast, over-expressing plants showed undeveloped adventitious roots, lateral roots, and a reduced root apical meristem. OsRPK1 over-expression also inhibited the expression of most auxin efflux carrier OsPIN genes, which was accompanied by changes in PAT and endogenous free IAA distribution in the leaves and roots.

Conclusions

The data indicated that OsRPK1, a novel leucine-rich-repeat receptor-like kinase, affects the root system architecture by negatively regulating polar auxin transport in rice.

General significance

This study demonstrated a common regulatory pathway of root system development in higher plants, which might be initiated by external stimuli via upstream receptor-like kinases and downstream carriers for polar auxin transport.     

Morphological and proteomic analysis of early stage of osteoblast differentiation in osteoblastic progenitor cells

Bone remodeling relies on a dynamic balance between bone formation and resorption, mediated by osteoblasts and osteoclasts, respectively. Under certain stimuli, osteoprogenitor cells may differentiate into premature osteoblasts and further into mature osteoblasts. This process is marked by increased alkaline phosphatase (ALP) activity and mineralized nodule formation. In this study, we induced osteoblast differentiation in mouse osteoprogenitor MC3T3-E1 cells and divided the process into three stages. In the first stage (day 3), the MC3T3-E1 cell under osteoblast differentiation did not express ALP or deposit a mineralized nodule. In the second stage, the MC3T3-E1 cell expressed ALP but did not form a mineralized nodule. In the third stage, the MC3T3-E1 cell had ALP activity and formed mineralized nodules. In the present study, we focused on morphological and proteomic changes of MC3T3-E1 cells in the early stage of osteoblast differentiation — a period when premature osteoblasts transform into mature osteoblasts. We found that mean cell area and mean stress fiber density were increased in this stage due to enhanced cell spreading and decreased cell proliferation. We further analyzed the proteins in the signaling pathway of regulation of the cytoskeleton using a proteomic approach and found upregulation of IQGAP1, gelsolin, moesin, radixin, and Cfl1. After analyzing the focal adhesion signaling pathway, we found the upregulation of FLNA, LAMA1, LAMA5, COL1A1, COL3A1, COL4A6, and COL5A2 as well as the downregulation of COL4A1, COL4A2, and COL4A4. In conclusion, the signaling pathway of regulation of the cytoskeleton and focal adhesion play critical roles in regulating cell spreading and actin skeleton formation in the early stage of osteoblast differentiation.     

粪产碱菌的Tn5转座诱变及吲哚乙酸生物合成特性的研究

粪产碱菌(Alcaligenes faecalis)A1501的吲哚乙酸(IAA)合成需要外源色氨酸参与。在不含色氨酸的限制性培养基中,A1501能良好生长,但不能合成IAA,表明在A1501中存在一条依赖于色氨酸的IAA合成途径。A1501的IAA合成具有菌体密度依赖特性。采用Tn5转座诱变技术构建A1501的突变库,从3500多株Tn5转染子中分离到一株色氨酸营养缺陷型突变株AT63。该Tn5突变株在不含色氨酸的限制性培养基上不能生长,但仍能进行IAA的生物合成,每毫升菌体密度等于10的突变株菌体的IAA合成量为224μg。对突变株AT63的研究表明在A1501中至少存在两条IAA合成途径：一条以色氨酸为合成前体,另一条以吲哚-3-磷酸甘油为前体。Southern杂交结果表明突变株中Tn5插入位点可能位于编码色氨酸合成酶基因上。     

Screening of Chromium-Resistant Bacteria for Plant Growth-Promoting Activities

Chromium-resistant plant growth-promoting bacteria (PGPB) were isolated from the electroplating industry waste disposal site soils at Coimbatore, India, using LB medium. The strain tolerated chromium concentrations up to 500 mg Cr⁶⁺L^?1 on LB medium. The strain was identified as Pseudomonas sp. VRK3 based on its morphological, physiological, and biochemical characteristics following Bergey's manual of determinative bacteriology. Evaluation of plant growth-promoting parameters revealed the intrinsic ability of the strain for the production of indole-3-acetic acid (IAA), siderophore, and solubilization of insoluble phosphate. Pseudomonas sp. VRK3 utilized tryptophan as a precursor for the growth and production of IAA (105.77 μg mL^?1) and also exhibited the production of siderophore. The strain utilized tricalcium phosphate as the sole source of phosphate exhibiting a high rate of phosphate solubilization (0.49 μg mL^?1). Pseudomonas sp. VRK3 showed a significant extent of chromium uptake and accumulation in their cell walls. Furthermore, Pseudomonas sp. VRK3 was demonstrated to possess mobilization of chromium from the soil that would enhance chromium availability to the plant. Potential use of this Pseudomonas sp. VRK3 as PGPB needs further testing in enhancing the growth and chromium uptake by the plants in pots under nursery conditions.     

四种叶菜衰老期间呼吸、乙烯产生、IAA和过氧化物酶的变化及其相互关系

结球白菜、结球甘蓝和菠菜在衰老期间的呼吸强度逐渐上升,后期下降,呈有峰型变化,而结球生菜的呼吸强度随衰老而下降,呈无峰型变化。外源乙烯使前三者的呼吸峰提前,但不改变其变化趋势,使后者出现呼吸峰。衰老期间内源乙烯均上升;过氧化物酶活力增加,IAA含量下降,两者存在极显著负相关。幼叶乙烯生成与IAA含量相关,老叶乙烯生成不受IAA水平影响。     

不同温度对青花菜花芽分化的影响

适当的低温有利于青花菜的花芽分化,14℃处理下的早熟青花菜植株可以完成花芽分化的各个过程,18℃处理下的植株花芽虽能分化,但花芽分化的各个过程不能完成,22℃处理下的植株一直处于营养生长状态而花芽未分化。叶中GA3和IAA含量伴随着花芽分化过程而变化,侧花茎上的每一级花芽分化过程中,GA3含量较高而IAA含量较低。     

表油菜素内酯促进小麦胚芽鞘伸长的作用

BR促进小麦胚芽鞘伸长的生理活性大于IAA,但高浓度的促进现象不如IAA明显。BR刺激乙烯生成与浓度相关。BR和IAA混合处理,对芽鞘切段的伸长、乙烯释放和H~+分泌都表现了加成作用。这二种激素在作用时间上有明显的差别,BR作用的滞后期更为清楚。BR有拮抗ABA对小麦胚芽鞘切段伸长的抑制作用。     

单性结实和自花结实柑橘果实发育过程中IAA、ZR和GA3含量的变化

对单性结实的龟井温州蜜柑(以下简称龟井)和自花结实的鄂柑1号橘(以下简称鄂柑1号)果实发育过程中不同部位的IAA、ZR和GA3的含量变化进行了测定。结果表明：(1)两品种果皮IAA含量呈相似的变化趋势,均于果实增大期间出现明显高峰,但鄂柑1号的IAA高峰值显著较高;二者果皮ZR含量动态却相反,增大期间龟井果皮ZR的含量相对较高且趋上升,而鄂柑1号却趋下降;二者果皮GA3含量均于增大期间出现类似的上升过程,之后均趋下降。(2)龟井果肉IAA、ZR和GA3含量前期均较高,此后均明显下降并居较低水平;而鄂柑1号果肉IAA和GA3均于增大期间出现明显的上升且含量明显高于龟井,而ZR含量与龟井一样呈下降趋势,但ZR的含量更低。(3)鄂柑1号种子的IAA、ZR和GA3在花后72d均相对较高,之后急剧回落至相对较低的稳定水平。     

两株生菜根际芽孢杆菌(Bacillus spp.)的分离与特性研究

【目的】从生菜根际土中筛选出2株具有多种生物学特性、促生和生防效果的芽孢杆菌。【方法】土样经过80°C高温处理,得到2株细菌。通过形态学、生理生化、16S rRNA和gyr B基因鉴定菌株。对其溶磷、合成IAA和嗜铁素能力及对植物病原真菌的拮抗作用进行测定。用2株细菌处理生菜种子,评价其促生效果。用菌株WXD 3-2处理小麦,评价其生防效果。【结果】经过鉴定,确定菌株WXD 3-1为巨大芽孢杆菌(Bacillus megaterium),WXD 3-2为枯草芽孢杆菌(Bacillus subtilis)。2株菌均有溶磷、合成嗜铁素、IAA能力和促生能力,菌株WXD 3-2能够对多种病原菌产生拮抗作用,抑制其生长。经过WXD 3-1和WXD 3-2处理,生菜植株高、叶片宽、植株鲜重及植株干重与对照相比分别增加21.51%和8.88%、31.93%和14.51%、41.30%和13.58%、42.76%和26.35%。菌株WXD 3-2能够减轻小麦根腐病病症,小麦根部病斑减少。【结论】分离出的2株芽孢杆菌均具有溶磷、合成IAA和嗜铁素能力,能够促进生菜的生长,且菌株WXD 3-2还具有生防效果。     

Identification of indole-3-butyric acid as an endogenous constituent of maize kernels and leaves

Indole-3-butyric acid (IBA) was identified by thin layer chromatography, gas-liquid chromatography and gas chromatography-mass spectrometry in kernels and leaves of corn (Zea mays) var. Hazera 224. Free and ester conjugated IBA were present in dry and germinating corn kernels and leaves. This is the first report of IBA in a monocotyledonous plant and, as far as we know, the first evidence for the presence of conjugated IBA.