Identification of a novel <Emphasis Type="Italic">SPLIT</Emphasis>-<Emphasis Type="Italic">HULL</Emphasis> (<Emphasis Type="Italic">SPH</Emphasis>) gene associated with hull splitting in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Key message

The split-hull phenotype caused by reduced lemma width and low lignin content is under control of SPH encoding a type-2 13-lipoxygenase and contributes to high dehulling efficiency.

Abstract

Rice hulls consist of two bract-like structures, the lemma and palea. The hull is an important organ that helps to protect seeds from environmental stress, determines seed shape, and ensures grain filling. Achieving optimal hull size and morphology is beneficial for seed development. We characterized the split-hull (sph) mutant in rice, which exhibits hull splitting in the interlocking part between lemma and palea and/or the folded part of the lemma during the grain filling stage. Morphological and chemical analysis revealed that reduction in the width of the lemma and lignin content of the hull in the sph mutant might be the cause of hull splitting. Genetic analysis indicated that the mutant phenotype was controlled by a single recessive gene, sph (Os04g0447100), which encodes a type-2 13-lipoxygenase. SPH knockout and knockdown transgenic plants displayed the same split-hull phenotype as in the mutant. The sph mutant showed significantly higher linoleic and linolenic acid (substrates of lipoxygenase) contents in spikelets compared to the wild type. It is probably due to the genetic defect of SPH and subsequent decrease in lipoxygenase activity. In dehulling experiment, the sph mutant showed high dehulling efficiency even by a weak tearing force in a dehulling machine. Collectively, the results provide a basis for understanding of the functional role of lipoxygenase in structure and maintenance of hulls, and would facilitate breeding of easy-dehulling rice.

     

Effect of silicon on grain yield of rice under cadmium-stress

Many publications indicated various beneficial effects of the addition of silicon (Si) in soil on the physiology of rice plants. The gene responsible for the Si-uptake in rice, low Si-influx 1 (Lsi1), was identified and cloned for this study. The photosynthetic rate (Pn), grain yield, and resistance to Cadmium (Cd)-stress of the wild-type (WT) and Lsi1-transgenic Lemont rice lines under Cd-stress were examined in an attempt to better understand the mechanism associated with the Si-addition, Cd-stress, and rice physiology. Si-fertilization significantly reduced the Cd-content in rice under Cd-stress. The effect was most significant in the Lsi1-overexpression transgenic Lemont rice (Lsi1-OE line) under high Cd-stress. Conversely, Cd in soil lowered the Si-uptake of the plants indicating a significant interaction between the two elements. During the grain-filling period, Cd-stress greatly reduced the chlorophyll content and Pn of the rice resulting in a diminished grain output. However, Lsi1-OE line with a higher chlorophyll content and Pn than either WT or Lsi1-RNAi transgenic Lemont rice (Lsi1-RNAi line) maintained a high photo-assimilate transportation for high yield potential. At harvest, Lsi1-OE line contained more Si and less Cd than WT, whereas the Lsi1-RNAi line showed an opposite result. In general, Cd-stress reduced, while Si-fertilization significantly increased, the grain yield on rice. However, no significant difference on the grain yields existed between WT and Lsi1-RNAi line. This might be due to a compensation effect generated by Lsi1-RNAi line. It appeared that Si in the soil, as well as the enhancing or inhibiting Lsi1 expression and the resistance to Cd-toxicity of the plants, could significantly affect the rice yield making alternations on these factors a plausible approach for production improvement.     

<Emphasis Type="Italic">Lsi1</Emphasis>-regulated Cd uptake and phytohormones accumulation in rice seedlings in presence of Si

Silicon (Si) is known for its role in regulating the response of plants to imposed abiotic stresses. Since the stresses generally hinder production of a crop, such as rice, the exploration of the biochemistry and plant physiology relating to the function is of interest. Indeed, recently, there were reports on the function of Lsi1 in regulating the tolerance of rice to cadmium (Cd) stress. This study compared the kinetics of the Cd uptakes in Lemont wild type rice and its transgenic lines exposed to Cd with or without exogenous Si supply. At the same time, changes on the endogenous phytohormones and growth of the rice seedlings were monitored. Genetically, Lsi1 overexpression was found to downregulate K_m and V_max of Cd uptake kinetics in the plants under Cd stress, especially in the presence of Si. On the other hand, Lsi1 RNAi upregulated K_m and V_max regardless whether Si was present or not. It implied that Lsi1 could be capable of regulating Si as well as Cd transports. Under Cd stress, addition of Si reduced the Cd uptake of the rice lines in the order of Lsi1-overexpression line?>?Lemont?>?Lsi1-RNAi line. In addition, it also affected the chlorophyll biosynthesis and dry mass accumulation of the rice plants under Cd stress. Analyses on phytohormones including IAA, GA₃, JA, SA and ABA, as well as physiological functions, of the seedlings further verified the active involvement of Lsi1 in the complex defense system of the plants against Cd stress.     

Physiological,biochemical, and molecular differences in chloroplast synthesis between leaf and corolla of cabbage (<Emphasis Type="Italic">Brassica rapa</Emphasis> L. var. chinensis) and rapeseed (<Emphasis Type="Italic">Brassica napus</Emphasis> L.)

Cadmium (Cd) pollution is one of the major concerns in the development of sustainable agriculture, particularly for rice production. Silicon (Si) was recently recognized for its ability to mitigate a variety of abiotic stresses including that caused by Cd on rice. However, mechanism of the complex process is still not fully understood. Under Cd-stress, the low Si-influx 1-RNAi transgenic Lemont rice (Lsi1-RNAi line) exhibited an increased Cd-uptake over its counterparts, the wild type and the Lsi1-overexpression transgenic rice (Lsi1-OE line). In contrast, the Lsi1 expression-enhanced Lsi1-OE line showed the greatest Si-uptake among the three lines, with the highest activities on anti-oxidants (such as, superoxide dismutase) and the lowest content of malondialdehyde. Lsi1 also displayed a negative regulation on Low Cd gene and the natural resistance-associated macrophage proteins, Nramp5, indicating its capacity to alleviate Cd-stress on rice. The results obtained by this study suggested that the mitigation of Cd-toxicity on rice by Si might involve functions, such as the inhibition on Cd-uptake and transport and the enhancement on anti-oxidative enzyme activities, as well as the Lsi1-related expression on regulation of Si-uptake in rice. A new avenue might become available for overcoming the rampant pollution that threatens the rice production in China.     

UV-B stress-induced ABA production in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> mutants defective in ethylene signal transduction pathway

In this study, we examined the influence of UV-B radiation (280–320 nm) on ABA accumulation in 14-day-old Arabidopsis thaliana (L.) Heynh plants of wild type (WT), ethylene receptor mutant (etr1-1), and mutant with a constitutively active ethylene signal transduction pathway (ctr1-1). ABA content in nonirradiated WT plants was twice higher than in each mutant. UV-B irradiation caused dose-dependent ABA accumulation in WT plants. In the etr1-1 mutant, the amount of accumulated ABA was significantly less. In the ctr1-1 mutant, ABA content didn’t increase after UV-B irradiation. These data suggest that start of stress-induced ABA formation requires the adjustable ethylene signal pathway. In the ctr1-1 mutant, a constitutively active (nonadjustable) ethylene signal pathway blocks stress-induced ABA accumulation.     

Content of Osmolytes and Flavonoids under Salt Stress in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> Plants Defective in Jasmonate Signaling

The effects of the salt stress (200 mM NaCl) and exogenous jasmonic acid (JA) on levels of osmolytes and flavonoids in leaves of four-week-old Arabidopsis thaliana L. plants of the wild-type (WT) Columbia-0 (Col-0) and the mutant jin1 (jasmonate insensitive 1) with impaired jasmonate signaling were studied. The increase in proline content caused by the salt stress was higher in the Col-0 plants than in the mutant jin1. This difference was especially marked if the plants had been pretreated with exogenous 0.1 μM JA. The sugar content increased in response to the salt stress in the JA-treated WT plants but decreased in the jin1 mutant. Treatment with JA of the WT plants but not mutant defective in jasmonate signaling also enhanced the levels of anthocyanins and flavonoids absorbed in UV-B range in leaves. The presence of JA increased salinity resistance of the Col-0 plants, since the accumulation of lipid peroxidation products and growth inhibition caused by NaCl were less pronounced. Under salt stress, JA almost did not render a positive effect on the jin1 plants. It is concluded that the protein JIN1/MYC2 is involved in control of protective systems under salt stress.     

Redescription of <Emphasis Type="Italic">Lagocephalus cheesemanii</Emphasis> (Clarke 1897), a senior synonym of <Emphasis Type="Italic">Lagocephalus gloveri</Emphasis> Abe and Tabeta 1983, based on morphological and genetic comparisons (Actinopterygii: Tetraodontiformes: Tetraodontidae)

Lagocephalus gloveri Abe and Tabeta 1983 is shown to be a junior synonym of Tetrodon cheesemanii Clarke 1897 based on examination of morphological characters and DNA analysis of specimens collected from the western North Pacific, Australia and New Zealand. Lagocephalus cheesemanii is distinguished from other species of Lagocephalus by the following combination of characters: spinules on the back in a rhomboidal patch extending from the region between the nasal organ to the posterior part of the pectoral fin; caudal fin double emarginate with middle rays posteriorly produced; dorsal-fin rays 11–15; anal-fin rays 11–14; pectoral-fin rays 15–18; vertebrae 8+11=19; dorsal half of the body dark brown to brownish black; caudal fin dark brown to black with dorsal and ventral white tips. A neotype of L. cheesemanii is designated.     

Characterization of the γ-aminobutyric acid shunt pathway and oxidative damage in Arabidopsis thaliana pop 2 mutants under various abiotic stresses

In the present study, three Arabidopsis thaliana pop2 mutant lines with different T-DNA insertions in a gene coding γ-aminobutyric acid transaminase (GABA-TA) were screened for seed germination percentage, stress-induced oxidative damage, and GABA content and metabolism under various abiotic stresses including high temperature (42 °C), low temperature (4 °C), salinity (NaCl), and osmotic stress (mannitol). All mutant lines showed a decreased germination under all the stress treatments with a significant reduction in the pop2-1 and pop2-3 mutant lines. Content of GABA and MDA increased significantly in all pop2 mutants and wild type (WT) seedlings in response to all the treatments. However, content of GABA and MDA was lower in all pop2 mutants comparing to the WT under the same treatments. GABA increased already after 30 min and increased significantly after 2 h at 42 °C especially in the pop2-3 and WT seedlings. In response to the cold treatment, GABA content increased up to 4-fold compared to the control in all pop2 mutants and WT seedlings. In response to the NaCl treatment, GABA accumulated slightly in the WT and all pop2 mutants. On the contrary, GABA content increased significantly in the pop2, pop2-1, and pop2-3 mutants and WT under all mannitol treatments.     

Comparison of rice flowering-time genes under paddy conditions

Heading date is one of most important agronomic traits in rice. Flowering regulatory mechanisms have been elucidated in many cultivars through various approaches. Although study about flowering has been extensively examined in rice, but contributions of floral regulators had been poorly understood in a common genetic background for rice grown under paddy conditions. Thus, we compared the expression of 10 flowering-time genes — OsMADS50, OsMADS51, OsVIL2, OsPhyA, OsPhyB, OsPhyC, Ghd7, Hd1, OsGI, and OsTrx1 — in the same genetic background for ‘Dongjin’ rice (Oryza sativa) grown under paddy conditions when days were longer than 13.5 h. Whereas the wild type (WT) rice flowered 105 days after sowing, the latest mutant to do so was ostrx1, flowering 53 d later. This indicated that the gene is the strongest inducer among all of those examined. Mutations in OsMADS50 delayed flowering by 45 d when compared with the WT, suggesting that this MADS gene is another strong positive element. The third positive element was OsVIL2; mutations in the gene caused plants to flower 27 d late. In contrast, the double phytochrome mutant osphyA osphyB flowered 44 d earlier than the WT. The single mutant osphyB and the double mutant osphyB osphyC did the same, although not as early as the osphyA osphyB double mutant. These results demonstrated that phytochromes are major inhibitors under paddy conditions. Mutations in Ghd7 accelerated flowering by 34 d, indicating that the gene is also a major inhibitor. The hd1 mutants flowered 16 d earlier than the WT while a mutation in OsGI hastened flowering by 10 d, suggesting that both are weak flowering repressors. Of the two florigen genes (Hd3a being the other one), RFT1 played a major role under paddy conditions. Its expression was strongly promoted by Ehd1, which was negatively controlled by Ghd7. Here we show that phytochromes strongly inhibit flowering and OsTrx1 and OsMADS50 significantly induce flowering under paddy conditions through Ghd7-Ehd1-RFT1 pathway. Thus, we may be able to control heading date under paddy conditions through manipulating those genes, Ghd7, Ehd1 and RFT1.     

Combined effect of ethylene- and salicylic acid-signaling insensitive mutation on Arabidopsis response to low temperature

The roles of ethylene (ET) or salicylic acid (SA) in plant response to low temperature (LT, 5 °C) have been implicated. However, the combined effect of ET- and SA-signaling on plant growth and metabolism under LT remains to be evaluated. In this study, we comparatively analyzed the response of Arabidopsis ethylene insensitive (ein) 2-1 (an ET insensitive mutant), nonexprressor of pathogenesis relative (npr)1-1 (an SA insensitive mutant) and double mutant ein2-1/npr1-1 plants to LT. The results show that a LT of 5 °C induced plant growth retardation to a less degree in ein2-1, an intermediate degree in npr1-1, but a much larger in ein2-1/npr1-1 compared to the wild-type (WT) plants. The LT susceptibility of the ein2-1/npr1-1 plants was correlated to a lower net photosynthetic rate and proline content, and a higher content of H₂O₂ and malondialdehyde and electrolyte leakage relative to the WT plants. Lower activities of superoxide dismutase, peroxidase, and catalase, as well as a lower glutathione content and a ratio of its reduced form to its oxidized form were also observed in the double mutant plants as compared with the WT plants. However, at normal conditions (23 °C), all the tested physiological and biochemical parameters were comparable between the ein2-1/npr1-1 and WT plants, and plant growth was even better in the double mutant than in the WT plants. On the contrary, most of the above-mentioned parameters were advantageous in the ein2-1 and npr1-1 plants over the WT plants under the LT conditions. These data suggest that a parallel function or physiological redundancy of nonexpressor of pathogenesis relative 1 and ethylene insensitive 2 existed in the Arabidopsis plant response to the LT. On the other hand, an interaction between ET- and SA-signaling occurred during this process.     

Prevalence of <Emphasis Type="Italic">VRN1</Emphasis> locus alleles among spring common wheat cultivars cultivated in Western Siberia

With the use of allele-specific primers developed for the VRN1 loci, the allelic diversity of the VRN-A1, VRN-B1, and VRN-D1 genes was studied in 148 spring common wheat cultivars cultivated under the conditions of western Siberia. It was demonstrated that modern Western Siberian cultivars have the VRN-A1a allele, which is widely distributed in the world (alone or in combination with the VRN-B1a and VRN-B1c alleles). It was established that the main contribution in acceleration of the seedling–heading time is determined by a dominant VRN-A1a allele, while the VRNA1b allele, on the contrary, determines later plant heading. Cultivars that have the VRN-A1b allele in the genotype are found with a frequency of 8%. It was shown that cultivars with different allele combinations of two dominant genes (VRN-A1a + VRN-B1c and VRN-A1a + VRN-B1a) are characterized by earlier heading and maturing.     

Brassinosteroid (BR) biosynthetic gene <Emphasis Type="Italic">lhdd10</Emphasis> controls late heading and plant height in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Key message

A Brd2 allele suppresses heading date by altering the expression of heading date regulators such as OsMADS50 , and also negatively regulates chlorophyll biosynthesis.

Abstract

Heading date and plant height are important determinants of yield in rice (Oryza sativa L.). In this study, we characterized a late heading, dwarf mutant known as lhdd10 selected following ethyl methane sulfonate (EMS)-treatment of ssp. indica cultivar 93-11. lhdd10 showed late heading, dwarfness and slightly darker-green leaves than wild-type 93-11 under long-day and short-day conditions. We isolated lhdd10 by map-based cloning; it encoded a putative FAD-linked oxidoreductase protein (a brassinosteroid biosynthetic gene) that localized to the nucleus. LHDD10 was constitutively expressed in various tissues, but more so in shoot apices and panicles. Our data showed that lhdd10 influences heading date by controlling the expression of heading date regulators, such as OsMADS50 in both LD and SD conditions. lhdd10 also negatively regulated expression of chlorophyll biosynthetic genes to reduce the chlorophyll content. Our data indicated that BRs play important roles in regulating heading date and chlorophyll biosynthesis. This work provides material that will allow study of how BRs regulate heading date in rice.

     

Three new species of <Emphasis Type="Italic">Spadicoides</Emphasis> from Lushan Mountain,China

Spadicoides lushanensis sp. nov., S. multiseptata sp. nov. and S. rostrata sp. nov. are described and illustrated from specimens collected on dead branches of unidentified plants in Lushan Mountain, China. Spadicoides lushanensis is characterized by obclavate, rostrate, pale brown, 40–65?×?4.5–5.5 μm, smooth, 6–8-euseptate conidia. Spadicoides multiseptata is easily distinguished by obclavate, smooth, 11–15-euseptate, 65–115?×?11.5–13 μm, brown to pale brown conidia with a subacute apex. Spadicoides rostrata differs from other described Spadicoides species by obclavate, rostrate, pale brown, 40–65?×?10.5–12.5 μm, smooth, predominantly 5-euseptate conidia. A dichotomous key to Spadicoides species is provided.     

Genetic complementation analysis of rice sucrose transporter genes in Arabidopsis <Emphasis Type="Italic">SUC2</Emphasis> mutant <Emphasis Type="Italic">atsuc2</Emphasis>

Sucrose transporters (SUTs) play a critical role on the phloem plasma membrane in loading sucrose into the phloem of source leaves for long-distance transport to sink organs. Rice has a small gene family of five SUTs, Oryza sativa SUT1 (OsSUT1) to OsSUT5. To identify rice SUTs that function as phloem loaders, we adopted a growth restoration assay of the severe growth retardation phenotype of atsuc2, a mutant of the best-characterized Arabidopsis phloem loader AtSUC2, by introducing OsSUTs. The rice SUT genes were expressed by two different promoters, the native phloem-specific promoter of AtSUC2 (pAtSUC2) and the constitutive Cauliflower Mosaic Virus 35S (pCaMV35S) promoter. Of all the transgenic atsuc2 plants, only pAtSUC2: OsSUT1 complemented the atsuc2 mutant phenotype in a comparable manner to wild type (WT), and consistent levels of soluble sugars and starch were recovered compared to those of WT. This suggests that OsSUT1 is a functional ortholog of the Arabidopsis AtSUC2 and functions as an apoplastic phloem loader. In addition, ossut1 mutants were produced via anther culture and their primary carbohydrate levels and growth phenotypes were indistinguishable from those of WT. This suggests that the rice phloem loader OsSUT1 function may not be essential for rice vegetative growth under normal conditions.     

RNA binding protein QKI contributes to WT1 mRNA and suppresses apoptosis in ST cells

The RNA binding protein quaking (QKI), a key member of the STAR family, as an upstream gene could involve in much process including cell proliferation, apoptosis, differentiation and so on. However, the roles of QKI in germ cell, especially in swine testis (ST) cells, was not clear currently. And apoptosis plays important roles in the growth and development. The purpose of the present study was to clarify the relationship between QKI and apoptosis in ST cells. Firstly, our results showed that pEF1α-QKI and shQKI3 have clear effects on expression levels of QKI. Secondly, we established that QKI directly binds to WT1 3′UTR by binding with QRE-1 (2046–2052 bp, ACTAAC) only. Furthermore, QKI overexpression significantly increased the expression levels of WT1 and Bcl-2. QKI also has the effect on delaying the degradation of WT1 mRNA. In addition, we verified that QKI had a significantly suppressed apoptosis in ST cells. Finally, pBI-WT1 could make up for shQKI3-induced decrease in WT1, Bcl-2 mRNA levels and suppress apoptosis in ST cells. The results demonstrated that QKI was an important regulatory factor that affects apoptosis by targeting WT1 gene.     

Overexpressing osa-miR171c decreases salt stress tolerance in rice

The miRNA171 family is one of the well-conserved miRNA families, and its role under stresses is not known except its expression on genome-wide expression analyses. osa-miR171c was induced by high concentration of salt (150 mM NaCl). A rice dh mutant with osa-miR171c overexpression triggered by a T-DNA insertion, significantly decreased salt tolerance at the stages of germination and seedling. This phenotype was confirmed by osa-miR171c overexpression transgenic rice. Compared with wild-type (WT), dh mutant reduced amounts of free proline and increased the water loss rate after salt treatment. Stomatal density in the leaf epidermis of dh mutant also increased. Moreover, dh mutant increased sensitivity to ABA treatment. Several stressresponsive genes were down-regulated in dh mutant than in WT under salty stress. These results indicate that osamiR171c is involved in modulating physiological changes, stomatal development, ABA-dependent pathways and expression of stress-related genes; thereby, it possibly contributes to salty tolerance.     

The stay-green phenotype of wheat mutant <Emphasis Type="Italic">tasg1</Emphasis> is associated with altered cytokinin metabolism

Key message

By measuring the cytokinin content directly and testing the sensitivity to the cytokinin inhibitor lovastatin, we demonstrated that tasg1 cytokinin metabolism is different from wild-type.

Abstract

Our previous studies have indicated that compared with wild-type (WT) plants, a wheat stay-green mutant tasg1 exhibited delayed senescence. In this study, we found that the root development of tasg1 occurred later than that of WT. The number of lateral roots was fewer, but the lateral root length was longer in tasg1 than in WT, which resulted in a lower root to shoot ratio in tasg1 than WT. The levels of cytokinin (CK), CK activity, and expression of CK metabolic genes were measured. We found that the total CK content in the root tips and leaf of tasg1 was greater than in WT. The accumulation of mRNA of the CK synthetic gene (TaIPT) in tasg1 was higher than in WT at 9 and 11 days during seedling growth, but the expression of CK oxidase gene (TaCKX) was significantly lower in tasg1. Furthermore, the CK inhibitor lovastatin was used to inhibit CK activity. When treated with lovastatin, both the chlorophyll content and thylakoid membrane protein stability were significantly lower in tasg1 than WT, consistent with the inhibited expression of senescence-associated genes (TaSAGs) in tasg1. Lovastatin treatment also inhibited the antioxidative capability of wheat seedlings, and tasg1 was more sensitive to lovastatin than WT, as indicated by the MDA content, protein carbonylation, and antioxidant enzyme activity. The decreased antioxidative capability after lovastatin treatment may be related to the down-regulation of some antioxidase genes. These results suggest that the CK metabolism was altered in tasg1, which may play an important role in its ability to delay senescence.