Physical Basis for Altered Stem Elongation Rates in Internode Length Mutants of Pisum

Biophysical parameters related to gibberellin (GA)-dependent stem elongation were examined in dark-grown stem-length genotypes of Pisum sativum L. The rate of internode expansion in these genotypes is altered due to recessive mutations which affect either the endogenous levels of, or response to, GA. The GA deficient dwarf L181 (ls), two GA insensitive semierectoides dwarfs NGB5865 and NGB5862 (Ika and Ikb, respectively) and the `slender' line L197 (la cry^[ill]), which is tall regardless of GA content, were compared to the wild-type tall cultivar, Torsdag. Osmotic pressure, estimated by vapor pressure osmometry, and turgor pressure, measured directly with a pressure probe, did not correlate with the differences in growth rate among the genotypes. Mechanical wall properties of frozen-thawed tissue were measured using a constant force assay. GA deficiency resulted in increased wall stiffness judged both on the basis of plastic compliance and plastic extensibility normalized for equal stem circumference. Plastic compliance was not reduced in the GA insensitive dwarfs, though Ika reduced circumference-normalized plasticity. In contrast, in vivo wall relaxation, determined by the pressure-block technique, differed among genotypes in a manner which did correlate with extension rates. The wall yield threshold was 1 bar or less in the tall lines, but ranged from 3 to 6 bars in the dwarf genotypes. The results with the ls mutant indicate that GA enhances stem elongation by both decreasing the wall yield threshold and increasing the wall yield coefficient. In the GA-insensitive mutants, Ika and Ikb, the wall yield threshold is substantially elevated. Plants possessing Ika may also possess a reduced wall yield coefficient.     

Contrasting Strategies of Alfalfa Stem Elongation in Response to Fall Dormancy in Early Growth Stage: The Tradeoff between Internode Length and Internode Number

Fall dormancy (FD) in alfalfa (Medicago sativa L.) can be described using 11 FD ratings, is widely used as an important indicator of stress resistance, productive performance and spring growth. However, the contrasting growth strategies in internode length and internode number in alfalfa cultivars with different FD rating are poorly understood. Here, a growth chamber study was conducted to investigate the effect of FD on plant height, aboveground biomass, internode length, and internode number in alfalfa individuals in the early growth stages. In order to simulate the alfalfa growth environment in the early stage, 11 alfalfa cultivars with FD ratings from one to 11 were chosen and seeded at the greenhouse, and then were transplanted into an artificial growth chamber. The experimental design was a randomized complete block in a split-plot arrangement with three replicates. Plant height, above-ground biomass, internode length, and internode number were measured in early growth stage in all individuals. Our findings showed that plant height and the aboveground biomass of alfalfa did not significantly differ among 11 different FD rated cultivars. Also, internode length and internode number positively affected plant height and the aboveground biomass of alfalfa individuals and the average internode length significantly increased with increasing FD rating. However, internode number tended to sharply decline when the FD rating increased. Moreover, there were no correlations, slightly negative correlations, and strongly negative correlations between internode length and internode number in alfalfa individuals among the three scales, including within-FD ratings, within-FD categories and inter-FD ratings, respectively. Therefore, our results highlighted that contrasting growth strategies in stem elongation were adopted by alfalfa with different FD ratings in the early growth stage. Alfalfa cultivars with a high FD rating have longer internodes, whereas more dormant alfalfa cultivars have a larger number of internodes. There were tradeoffs between internode length and internode number in response to FD in alfalfa, which reflected certain scale-dependence.     

The Role of Farnesol as a Regulator of Stomatal Opening in Sorghum

Fine, very dilute aqueous emulsions of all-trans farnesol appliedto intact leaves of Sorghum bicolor caused appreciable inhibitionof stomatal opening which persisted for 2 d, after which timethe stomata regained their capacity to open. The inhibitoryeffect of farnesol was not overcome by flushing the leaves withCO₂-free air, indicating that it was not the result of an accumulationof CO₂. This conclusion was supported by measurements of CO₂compensation, which increased only slightly after farnesol treatment. All-trans farnesol has previously been reported to be formedin water-stressed plants of Sorghum. The data presented heresuggest that it could be acting as an endogenous antitranspirant,in a comparable role to that already established for abscisicacid in several species. It would appear, however, to have aless prolonged inhibitory effect than abscisic acid, and itcould be responsible for the rapid responses of Sorghum stomatato water stress and their quick recovery after the plant hasregained turgor, a characteristic which distinguishes Sorghumfrom many other genera so far investigated.     

Internode Length in Pisum: Variation in Response to a Daylength Extension with Incandescent Light

Lines representing a range of internode length and floweringgenotypes in Pisum sativum L. were grown in 8 h of daylightfollowed by either 16 h of darkness or incandescent light. Thestem elongation response index (RI = length in 24 h ÷length in 8 h) was least in the very short internode nana types,which are grossly deficient in gibberellins (GAs), and the verylong internode slender types, which behave as if saturated withGAs. The common tall (genotype Le) and dwarf (le) types (lepartially blocks conversion of GA₂₀ to the active form, GA₁)were all markedly responsive but the peak RI (based on the mostresponsive internode) was less in tall lines (1.79 to 2.78)than in dwarf lines (2.32 to 5.01) and the peak RI tended tooccur about three to four internodes earlier in tall than indwarf lines. The cry⁸ mutation reduced the RI. (Duplicate lengthloci La and Cry are probably concerned with GA reception.) Amongle dwarf lines, genotype La cry⁸, was generally less responsivethan La Cry, La cry^c and la Cry. Data from crosses showed thaton either an le La or le la background cry⁸ segregates had alower RI than cry⁸ segregates. On an le la background, cry⁸plants were shorter than cry^c plants, cry⁸ was partially dominantto cry⁸ and segregation was clear only in long days. On an lela background, cry^c plants were shorter than cry^c plants, cry⁸was partially dominant to cry⁸ and segregation was clear inlong or short days. The very high peak RI (5.0) of the microcryptodwarfline, L57, appeared to result, in part, from a marked foreshorteningof internodes 4 to 10 in the 8 h regime. In the 24 h regimeL57 (lm) had a fairly similar growth pattern to normal (Lm)cryptodwarf types. The peak RI tended to occur at a lower internode in early thanlate flowering lines, especially among dwarf types, and genotypeswith a day neutral flowering habit (genotype sn or dne) wereless responsive than their photoperiodic counterparts (Sn Dne). White fluorescent light, given as a daylength extension, wasmuch less effective than incandescent light at stimulating stemelongation suggesting control through the phytochrome equilibrium(P_tr/P_total). Pisum sativum, garden pea, daylength extension, flowering, genotype, gibberellin, hormone receptor, incandescent light, internode length, phytochrome, stem elongation     

Identification of rCop-1, a New Member of the CCN Protein Family,as a Negative Regulator for Cell Transformation

By using a model system for cell transformation mediated by the cooperation of the activated H-ras oncogene and the inactivated p53 tumor suppressor gene, rCop-1 was identified by mRNA differential display as a gene whose expression became lost after cell transformation. Homology analysis indicates that rCop-1 belongs to an emerging cysteine-rich growth regulator family called CCN, which includes connective-tissue growth factor, CYR61, CEF10 (v-src inducible), and the product of the nov proto-oncogene. Unlike the other members of the CCN gene family, rCop-1 is not an immediate-early gene, it lacks the conserved C-terminal domain which was shown to confer both growth-stimulating and heparin-binding activities, and its expression is lost in cells transformed by a variety of mechanisms. Ectopic expression of rCop-1 by retroviral gene transfers led to cell death in a transformation-specific manner. These results suggest that rCop-1 represents a new class of CCN family proteins that have functions opposing those of the previously identified members.Oncogenic conversion of a normal cell into a tumor cell requires multiple genetic alterations (12). Of particular interest is the fact that mutations in both ras oncogenes (3) and the p53 tumor suppressor gene cooperate in transformation of mammalian cells (11). Mutations in both ras and the p53 gene were also found at high frequencies in a variety of human cancers, including those of the colon, lung, and pancreas (2, 18). It has been proposed that both p53 and Ras function, whether directly or through other signaling molecules, to control expression of genes that are important for cell growth and differentiation (13, 17, 37). To this end, several ras target genes (10) and p53 target genes, including those encoding p21/CIP1/WAF1, an inhibitor of G₁ cyclin-dependent kinase (9); Mdm-2, a negative regulator of p53 (1); GADD45, a protein involved in DNA repair (36); and Bax, which promotes apoptosis (28), have been identified. Most of these genes, except p21/CIP1/WAF1, which was cloned by subtractive hybridization, were identified by the candidate gene hypothesis. Recently, more p53 target genes have been isolated by the differential display technique, including those coding for cyclin G (31); MAP4, a microtubule-associated protein negatively regulated by p53 (29); and PAG608, a novel nuclear zinc finger protein whose overexpression promotes apoptosis (14). Functional characterizations of these genes have shed light on the role of p53 in cell cycle control and apoptosis. However, genes that mediate tumor suppression activity by p53 remain elusive.The fact that neither the inactivation of p53 nor the activation of Ras alone is able to transform primary mammalian cells (34), whereas both mutations together can do so, suggests that genes regulated by p53 and Ras cooperate in upsetting normal cell growth control cells (11). Using differential display (22), we set out to identify genes whose expression is altered by both mutant ras and p53 by comparing the mRNA expression profiles of normal rat embryo fibroblasts (REFs) and their derivatives transformed by either a constitutively inactivated or a temperature-sensitive mutant p53 in cooperation with the activated H-ras oncogene (11, 27). In this report we describe the identification and give a functional characterization of rCop-1, a gene whose expression is abolished by cell transformation. By sequence homology, rCop-1 was found to belong to an emerging cysteine-rich growth regulator family called CCN (which stands for connective-tissue growth factor [CTGF], CEF10/Cyr61, and Nov) (4). Here we show that rCop-1 may represent a novel class of CCN family proteins based on its unique cell cycle expression pattern, its lack of the C-terminal (CT) domain conserved in all CCN proteins, its loss of expression in all transformed cells analyzed, and its ability to confer cytotoxicity to the transformed cells.     

Cell Walls and the Developmental Anatomy of the Brachypodium distachyon Stem Internode

While many aspects of plant cell wall polymer structure are known, their spatial and temporal distribution within the stem are not well understood. Here, we studied vascular system and fiber development, which has implication for both biofuel feedstock conversion efficiency and crop yield. The subject of this study, Brachypodium distachyon, has emerged as a grass model for food and energy crop research. Here, we conducted our investigation using B. distachyon by applying various histological approaches and Fourier transform infrared spectroscopy to the stem internode from three key developmental stages. While vascular bundle size and number did not change over time, the size of the interfascicular region increased dramatically, as did cell wall thickness. We also describe internal stem internode anatomy and demonstrate that lignin deposition continues after crystalline cellulose and xylan accumulation ceases. The vascular bundle anatomy of B. distachyon appears to be highly similar to domesticated grasses. While the arrangement of bundles within the stem is highly variable across grasses, B. distachyon appears to be a suitable model for the rind of large C₄ grass crops. A better understanding of growth and various anatomical and cell wall features of B. distachyon will further our understanding of plant biomass accumulation processes.     

Sar1, a Novel Regulator of ER-Mitochondrial Contact Sites

Endoplasmic reticulum (ER)—mitochondrial contact sites play a pivotal role in exchange of lipids and ions between the two organelles. How size and function of these contact sites are regulated remains elusive. Here we report a previously unanticipated, but conserved role of the small GTPase Sar1 in the regulation of ER-mitochondrial contact site size. Activated Sar1 introduces membrane curvature through its N-terminal amphiphatic helix at the ER-mitochondria interphase and thereby reducing contact size. Conversely, the S. cerevisiae N3-Sar1 mutant, in which curvature induction is decreased, caused an increase in ER-mitochondrial contacts. As a consequence, ER tubules are no longer able to mark the prospective scission site on mitochondria, thereby impairing mitochondrial dynamics. Consistently, blocking mitochondrial fusion partially rescued, whereas deletion of the dynamin-like protein enhanced the phenotype in the sar1D32G mutant. We conclude that Sar1 regulates the size of ER-mitochondria contact sites through its effects on membrane curvature.     

Polycomb Group Protein YY1 Is an Essential Regulator of Hematopoietic Stem Cell Quiescence

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Internode Length in Pisum: the Response of le na Scions Grafted to Na Stocks

The recessive gene na results in peas with extremely short internodes(phenotype nana). In intact plants, na prevents expression ofthe Le/le gene pair which determine the tall/dwarf difference.na blocks a step early in the gibberellin biosynthetic pathwaywhile le prevents conversion of gibberellin A₂₀ to GA₁. Whengrafted to leafy Na stocks, le na and Le na scions become phenotypicallydwarf and tall, respectively. Hence, Na stocks provide a graft-transmissiblesubstance which promotes elongation of na scions and allowsexpression of the Le/le difference. Pisum, internode length, grafting, genotype     

Inhibition of Anthocyanin Synthesis by Cobaltous Ions in the First Internode of Sorghum bicolor L. Moench

Cobaltous ions (Co²⁺) inhibited light-mediated anthocyanin synthesisin excised first internodes of Sorghum bicolor (L.) Moench.Studies with precursors/intermediates of the anthocyanin biosyntheticpathway, such as acetate, phenylalanine, t-cinnamic acid, tyrosine,and p-coumaric acid were undertaken to identify the metabolicsite at which Co²⁺ ions inhibit anthocyanin synthesis. p-Coumaricacid partially restored anthocyanin synthesis. No other intermediatewas effective in bringing about recovery. It is suggested thatCo²⁺ might interfere with process/es which leads to the formationof p-coumaric acid, an intermediate of the anthocyanin biosyntheticpathway. Key words: Sorghum bicolor (L.) Moench, anthocyanin, cobaltous ions, phenylalanine ammonia-lyase     

长穗颈温敏核不育水稻穗颈节间长度及细胞学观察

以水稻培矮64S作对照,对隐性长穗颈温敏核不育水稻(Oryza sativa L.)长选3S幼穗各发育时期穗颈节间长度及细胞数量和长度进行比较分析。结果表明,长选3S在幼穗分化的二核期至始花期穗颈节间伸长速度最快,其节间伸长的长度是培矮64S的2.1倍;在花粉母细胞减数分裂期至二核期及始花当天至始花后第3天两个时段,穗颈节间伸长速度慢。穗颈节间的细胞个数和平均长度两者呈现相似的变化规律,但从花粉母细胞减数分裂期至始花期,长选3S穗颈节间薄壁细胞增加3759个,细胞平均长度为72.9μm,培矮64S增加3134个,细胞平均长度为38μm,长选3S穗颈节间的伸长主要是由幼穗分化的花粉母细胞减数分裂期至始花期细胞分裂和细胞伸长共同作用的结果,其中后者的作用更显著。     

Increase in Internode Length of Phaseolus lunatus L. Caused by Inoculation with a Nitrate Reductase-deficient Strain of Rhizobium sp

Dramatic differences in the height of lima beans (Phaseolus lunatus L.) treated with two different Rhizobium strains were studied. Lima beans were grown in Perlite in the greenhouse or in a minus-N culture solution in the growth chamber. The plants were inoculated with either Rhizobium sp. (lima bean) strain 127E15, which contains the constitutive nitrate reductase activity, or strain 127E14, which lacks that activity. For up to 3 weeks, no growth differences were observed in the plants inoculated with either strain. Five weeks after inoculation, however, those plants inoculated with strain 127E14 were significantly taller and had a larger number of leaves than those inoculated with strain 127E15. The difference in plant height was the result of increased internode elongation caused by inoculation with Rhizobium sp. 127E14. This response was observed with all lima bean cultivars tested, including Henderson, Fordhook, Allgreen, and Early Thorogreen. The growth difference occurred in plants cultured in the greenhouse or in the growth chamber.     

Identification of a Novel Lipoprotein Regulator of Clostridium difficile Spore Germination

Clostridium difficile is a Gram-positive spore-forming pathogen and a leading cause of nosocomial diarrhea. C. difficile infections are transmitted when ingested spores germinate in the gastrointestinal tract and transform into vegetative cells. Germination begins when the germinant receptor CspC detects bile salts in the gut. CspC is a subtilisin-like serine pseudoprotease that activates the related CspB serine protease through an unknown mechanism. Activated CspB cleaves the pro-SleC zymogen, which allows the activated SleC cortex hydrolase to degrade the protective cortex layer. While these regulators are essential for C. difficile spores to outgrow and form toxin-secreting vegetative cells, the mechanisms controlling their function have only been partially characterized. In this study, we identify the lipoprotein GerS as a novel regulator of C. difficile spore germination using targeted mutagenesis. A gerS mutant has a severe germination defect and fails to degrade cortex even though it processes SleC at wildtype levels. Using complementation analyses, we demonstrate that GerS secretion, but not lipidation, is necessary for GerS to activate SleC. Importantly, loss of GerS attenuates the virulence of C. difficile in a hamster model of infection. Since GerS appears to be conserved exclusively in related Peptostreptococcaeace family members, our results contribute to a growing body of work indicating that C. difficile has evolved distinct mechanisms for controlling the exit from dormancy relative to B. subtilis and other spore-forming organisms.