RNA结合蛋白RB47、RB60在叶绿体翻译调节中的作用

光调节的叶绿体mRNA的翻译需要mRNA5′,非翻译区(5′ UTR)相互作用的中间作用因子.叶绿体多聚腺苷酸结合蛋白(cPABP)RB47特异性地与PsbAmRNA的5′ UTR结合,并对于该mRNA的翻译是很重要的.定位于叶绿体中与cPABP伴随纯化出来的一种蛋白质二硫化物异钩酶RB60表现出调节cPABP与PsbAmRNA的5′ UTR结合的特性.这种调节是通过可逆地改变有氧化还原特征的cPABP的氧化还原状态来实现,或通过ADP依赖的磷酸化来实现.这种机制就在调节叶绿体的基因表达中提供了一个简单可逆的开关机制.     

Inhibition of oxygen evolution by zaluzanin C

The inhibition of ATP synthesis, proton uptake, and electron transport (basal, phosphorylating, and uncoupled) from water to methylviologen by zaluzanin C indicates that it acts as an electron transport inhibitor. Uncoupled photosystem I electron transport rate was not affected by zaluzanin C; however, uncoupled photosystem II electron flow from water to 2,3,5,6-tetramethyl phenylenediamine and from water to silicomolybdate was inhibited, but electron transport from diphenylcarbazide to silicomolybdate was not affected. We conclude that the site of inhibition by zaluzanin C is located at the oxygen evolution level.     

The Role of E3 Ubiquitin Ligases in Chloroplast Function

Chloroplasts are ancient organelles responsible for photosynthesis and various biosynthetic functions essential to most life on Earth. Many of these functions require tightly controlled regulatory processes to maintain homeostasis at the protein level. One such regulatory mechanism is the ubiquitin-proteasome system whose fundamental role is increasingly emerging in chloroplasts. In particular, the role of E3 ubiquitin ligases as determinants in the ubiquitination and degradation of specific intra-chloroplast proteins. Here, we highlight recent advances in understanding the roles of plant E3 ubiquitin ligases SP1, COP1, PUB4, CHIP, and TT3.1 as well as the ubiquitin-dependent segregase CDC48 in chloroplast function.     

The Plastidial DIG5 Protein Affects Lateral Root Development by Regulating Flavonoid Biosynthesis and Auxin Transport in Arabidopsis

To reveal the mechanisms underlying root adaptation to drought stress, we isolated and characterized an Arabidopsis mutant, dig5 (drought inhibition of lateral root growth 5), which exhibited increased sensitivity to the phytohormone abscisic acid (ABA) for the inhibition of lateral root growth. The dig5 mutant also had fewer lateral roots under normal conditions and the aerial parts were yellowish with a lower level of chlorophylls. The mutant seedlings also displayed phenotypes indicative of impaired auxin transport, such as abnormal root curling, leaf venation defects, absence of apical hook formation, and reduced hypocotyl elongation in darkness. Auxin transport assays with [³H]-labeled indole acetic acid (IAA) confirmed that dig5 roots were impaired in polar auxin transport. Map-based cloning and complementation assays indicated that the DIG5 locus encodes a chloroplast-localized tRNA adenosine deaminase arginine (TADA) that is involved in chloroplast protein translation. The levels of flavonoids, which are naturally occurring auxin transport inhibitors in plants, were significantly higher in dig5 roots than in the wild type roots. Further investigation showed that flavonoid biosynthetic genes were upregulated in dig5. Introduction of the flavonoid biosynthetic mutation transparent testa 4 (tt4) into dig5 restored the lateral root growth of dig5. Our study uncovers an important role of DIG5/TADA in retrogradely controlling flavonoid biosynthesis and lateral root development. We suggest that the DIG5-related signaling pathways, triggered likely by drought-induced chlorophyll breakdown and leaf senescence, may potentially help the plants to adapt to drought stress through optimizing the root system architecture.     

采后小白菜叶绿体色素含量变化及其叶绿素降解动力学的研究

目的 探究采后小白菜贮藏过程中叶绿体色素的代谢变化及其叶绿素降解的动力学模型。方法 以采后“上海青”小白菜为试材, 分别将其贮藏于室温(20 ℃)、低温(2 ℃)及冰温(-0.5 ℃)条件下, 研究其总叶绿素、叶绿素a、叶绿素b、β-胡萝卜素及叶黄素的代谢变化, 并对其叶绿素的降解进行动力学模型拟合。结果 采后小白菜贮藏过程中总叶绿素、叶绿素a、叶绿素b、β-胡萝卜素和叶黄素均发生不同程度的降解, 低温和冰温贮藏均可减缓其降解, 其中冰温贮藏减缓效果最佳。叶绿素a含量与叶绿素b含量的比值、β-胡萝卜素含量与总叶绿素含量的比值以及室温贮藏条件下叶黄素含量与总叶绿素含量的比值均呈现不同程度的上升趋势, 而低温和冰温贮藏环境中的叶黄素含量与总叶绿素含量的比值总体则呈现下降趋势, 但贮藏6 d后下降不显著。此外, 叶绿素的降解反应符合零级动力学反应模型, 拟合调整决定系数(Adj.R2)均大于0.93, 降解活化能为95.53 kJ/mol, 降解动力学模型为k=6.51×1015?exp(9.553×104/RT)。结论 采后小白菜贮藏过程中叶绿体色素的代谢变化受贮藏温度影响较大, 其叶绿素降解反应符合零级动力学反应模型。     

The Plastome Sequences of Triticum sphaerococcum (ABD) and Triticum turgidum subsp. durum (AB) Exhibit Evolutionary Changes,Structural Characterization,Comparative Analysis,Phylogenomics and Time Divergence

The mechanism and course of Triticum plastome evolution is currently unknown; thus, it remains unclear how Triticum plastomes evolved during recent polyploidization. Here, we report the complete plastomes of two polyploid wheat species, Triticum sphaerococcum (AABBDD) and Triticum turgidum subsp. durum (AABB), and compare them with 19 available and complete Triticum plastomes to create the first map of genomic structural variation. Both T. sphaerococcum and T. turgidum subsp. durum plastomes were found to have a quadripartite structure, with plastome lengths of 134,531 bp and 134,015 bp, respectively. Furthermore, diploid (AA), tetraploid (AB, AG) and hexaploid (ABD, AGA^m) Triticum species plastomes displayed a conserved gene content and commonly harbored an identical set of annotated unique genes. Overall, there was a positive correlation between the number of repeats and plastome size. In all plastomes, the number of tandem repeats was higher than the number of palindromic and forward repeats. We constructed a Triticum phylogeny based on the complete plastomes and 42 shared genes from 71 plastomes. We estimated the divergence of Hordeum vulgare from wheat around 11.04–11.9 million years ago (mya) using a well-resolved plastome tree. Similarly, Sitopsis species diverged 2.8–2.9 mya before Triticum urartu (AA) and Triticum monococcum (AA). Aegilops speltoides was shown to be the maternal donor of polyploid wheat genomes and diverged ~0.2–0.9 mya. The phylogeny and divergence time estimates presented here can act as a reference framework for future studies of Triticum evolution.     

Chloroplast Thylakoidal Ascorbate Peroxidase,PtotAPX, Has Enhanced Resistance to Oxidative Stress in Populus tomentosa

Chloroplasts are the most major producers of reactive oxygen species (ROS) during photosynthesis. However, the function of thylakoid ascorbate peroxidase (tAPX) in response to oxidative stress in wood trees is largely unknown. Our results showed that PtotAPX of Populus tomentosa could effectively utilize ascorbic acid (AsA) to hydrolyze hydrogen peroxide (H₂O₂) in vitro. The overexpression or antisense of PtotAPX (OX-PtotAPX or anti-PtotAPX, respectively) in Populus tomentosa plants did not significantly affect plant morphology during plant growth. When treated with methyl viologen (MV), the OX-PtotAPX plants exhibited less morphological damage under stress conditions compared to WT plants. OX-PtotAPX plants maintained lower H₂O₂ levels and malondialdehyde (MDA) contents, but more reduced AsA levels, a higher photosynthetic rate (Pn), and the maximal photochemical efficiency of PSII (Fv/Fm), whereas anti-PtotAPX plants showed the opposite phenotype. Furthermore, the activity of APX was slightly higher in OX-PtotAPX under normal growth conditions, and this activity significantly decreased after stress treatment, which was the lowest in anti-P. Based on these results, we propose that PtotAPX is important for protecting the photosynthetic machinery under severe oxidative stress conditions in P. tomentosa, and is a potential genetic resource for regulating the stress tolerance of woody plants.     

Plastid Transformation of Micro-Tom Tomato with a Hemipteran Double-Stranded RNA Results in RNA Interference in Multiple Insect Species

Plant-mediated RNA interference (RNAi) holds great promise for insect pest control, as plants can be transformed to produce double-stranded RNA (dsRNA) to selectively down-regulate insect genes essential for survival. For optimum potency, dsRNA can be produced in plant plastids, enabling the accumulation of unprocessed dsRNAs. However, the relative effectiveness of this strategy in inducing an RNAi response in insects using different feeding mechanisms is understudied. To investigate this, we first tested an in vitro-synthesized 189 bp dsRNA matching a highly conserved region of the v-ATPaseA gene from cotton mealybug (Phenacoccus solenopsis) on three insect species from two different orders that use leaf-chewing, lacerate-and-flush, or sap-sucking mechanisms to feed, and showed that the dsRNA significantly down-regulated the target gene. We then developed transplastomic Micro-tom tomato plants to produce the dsRNA in plant plastids and showed that the dsRNA is produced in leaf, flower, green fruit, red fruit, and roots, with the highest dsRNA levels found in the leaf. The plastid-produced dsRNA induced a significant gene down-regulation in insects using leaf-chewing and lacerate-and-flush feeding mechanisms, while sap-sucking insects were unaffected. Our results suggest that plastid-produced dsRNA can be used to control leaf-chewing and lacerate-and-flush feeding insects, but may not be useful for sap-sucking insects.