An open-flower mutant of Melilotus alba: Potential for floral-dip transformation of a papilionoid legume with a short life cycle?

This paper describes an open-flower mutant, designated opf, that we discovered in a genetic screen of fast neutron bombardment mutants in an attempt at floral-dip transformation of Melilotus alba (Fabaceae; white sweetclover), an alternative papilionoid legume host for Sinorhizobium meliloti. The opf mutant developed flowers with reflexed sepals and petals, thereby exposing the stamens and carpel, whereas wild-type sweetclover inflorescences developed closed flowers where the young stamens and carpel remain covered during the early stages of flower development. Based on crosses with the wild type, the mutant segregated as a single, Mendelian recessive. Crosses were successful only when the opf mutant served as the female parent, suggesting that the mutant was male sterile. However, no obvious differences from wild-type stamen development were observed in the opf mutant. The anther defect was due to indehiscence. However, as the plants approached the end of their life cycle, the frequency of selfing increased. We also investigated whether the opf mutant could be transformed via Agrobacterium tumefaciens floral-dip infiltration because open flowers like those of Arabidopsis appear to be more readily transformable. However, similar to wild-type M. alba, the opf mutant is refractory to floral-dip transformation by Agrobacterium tumefaciens.     

Salt sensitivity of the vegetative and reproductive stages in chickpea (Cicer arietinum L.): Podding is a particularly sensitive stage

Soil salinity is an increasing problem, including in regions of the world where chickpea is cultivated. Salt sensitivity of chickpea was evaluated at both the vegetative and reproductive phase. Root-zone salinity treatments of 0, 20, 40 and 60 mM NaCl in aerated nutrient solution were applied to seedlings or to older plants at the time of flower bud initiation. Even the reputedly tolerant cultivar JG11 was sensitive to salinity. Plants exposed to 60 mM NaCl since seedlings, died by 52 d without producing any pods; at 40 mM NaCl plants died by 75 d with few pods formed; and at 20 mM NaCl plants had 78-82% dry mass of controls, with slightly higher flower numbers but 33% less pods. Shoot Cl exceeded shoot Na by 2-5 times in both the vegetative and reproductive phase, and these ions also entered the flowers. Conversion of flowers into pods was sensitive to NaCl. Pollen from salinized plants was viable, but addition of 40 mM NaCl to an in vitro medium severely reduced pollen germination and tube growth. Plants recovered when NaCl was removed at flower bud initiation, adding new vegetative growth and forming flowers, pods and seeds. Our results demonstrate that chickpea is sensitive to salinity at both the vegetative and reproductive phase, with pod formation being particularly sensitive. Thus, future evaluations of salt tolerance in chickpea need to be conducted at both the vegetative and reproductive stages.     

COMPARISON OF TETRAPLOID AND SINGLE GENE-INDUCED GIGAS VARIANTS IN CHICKPEA (CICER ARIETINUM). I. ORIGIN AND GENETIC CHARACTERIZATION

Two genetic variants with increased organ size were independently derived from diploid (2n = 2x = 16) chickpea (Cicer arietinum L.) line ICC 640. Radiation-induced mutant PM1101 had greatly enlarged leaves, leaflets, and pods, and an elongated stem with longer internodes but fewer nodes than ICC 640. F₁, F₂, and F₃ data from crosses with ICC 640 showed that the mutant characteristics of PM1101 were the pleiotropic effects of a single, recessive genetic factor. For purposes of comparison, tetraploid derivatives of ICC 640 were produced by colchicine treatment of seed. In the tetraploids, leaflets and pods were enlarged, but less dramatically than in PM1101. Enlarged pollen grains and stomatal guard cells, and increased guard cell chloroplast number were found in tetraploids but not in PM1101, while both variants produced fewer seeds than ICC 640. Mutant PM1101 and the tetraploids represent two very different manifestations of gigantism in chickpea.     

Growth and metabolic responses of contrasting chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L.) genotypes to chilling stress at reproductive phase

Chilling stress (<10°C) at reproductive phase of chickpea results in abortion of flowers and pods leading to poor yield. The metabolic causes associated with cold sensitivity of chickpea are not well understood. Hence, in the present study, we evaluated four chickpea genotypes (ICC 16348, ICC 16349, PBG1 and GPF2) having contrasting cold sensitivity for their reproductive growth and metabolism subjected to cold stress (average day temperature: 17.6°C; average night temperature: 4.9°C). Genotypes ICC 16348 and ICC 16349 showed flowering and set pods, while PBG1 and GPF2 failed to do so during the stress conditions indicating the former to be cold tolerant. The stress injury in the leaves such as increase in electrolyte leakage, decrease in chlorophyll content and relative leaf water content was significantly less in ICC 16348 and ICC 16349 genotypes. The analysis of carbohydrates indicated total sugars and starch to be present in greater content in ICC 16348 and ICC 16349 relative to PBG1 and GPF2 genotypes. The enzymes related to carbohydrate metabolism such as β-amylase, invertase and sucrose synthase showed significantly higher activity in the leaves of ICC 16348 and ICC 16349 compared to the other two genotypes. PBG1 and GPF2 genotypes experienced greater oxidative stress measured as malondialdehyde and hydrogen peroxide. ICCV 16348 and ICC 16349 possessed significantly higher levels of enzymatic (superoxide dismutase, catalase, ascorbate peroxidase) and non-enzymatic antioxidants (proline and ascorbic acid) relative to PBG1 and GPF2. Particularly, proline and ascorbic acid were markedly higher in cold-tolerant genotypes compared to the sensitive ones suggesting their deciding role in governing the cold tolerance.     

航天诱变百脉根结瘤突变体的筛选

通过返回式卫星搭载,利用太空环境对百脉根(Lotus japonicus)MG-20种子进行诱变。从种植的三代植株中,筛选到多种共生固氮根瘤的突变体,其中不结根瘤突变体18个株系,表现为接种根瘤菌两周后无根瘤形成;结无效根瘤突变体9个株系,表现为根瘤数目少且分布不均匀,根瘤呈白色,有些为半透明;花叶形态异常突变体1个株系,表现为除根瘤数目少外,植株矮小、托叶消失、花形态异常;纤细突变体1个株系,表现为除根瘤数目少外,植株变小、茎细叶小。     

大豆花荚败育期间的植物激素变化

大豆开花结荚期,不同发育阶段的幼蕾与花荚的脱落率不同,其中以花后5d内的幼荚脱落最严重。与败育花荚相比,正常花荚中的干物质积累量均较高。细胞分裂素(DHZRs,ZRs,iPA)含量也较高,花后3～5d的幼荚中表现更明显。脱落酸(ABA)则是以败育幼蕾及花后3～5d的幼荚中含量较高。不同发育阶段的大豆生殖器官中,正常开放花中的玉米赤霉烯酮(ZEN)含量最高。     

Inheritance of flower color in chickpea

Flower color is a useful morphological marker in chickpea (Cicer arietinum L.). Inheritance of this trait was studied using two white-flowered chickpea genotypes, P 9623 and RS 11, and one blue-flowered genotype, T 39-1. The genetic constitutions of the white flower colors of P 9623 and RS 11 were different, for in an earlier study their F1 produced pink flowers. The two F1s of the crosses P 9623 x T 39-1 and RS 11 x T 39-1 also produced pink flowers. Each of the two F2 populations segregated in 9 pink:3 blue:4 white-flowered plants. These results can be explained by a three-gene model. These three independently segregating genes are probably the same as C, B, and P reported in the literature earlier. Allelic tests could not be undertaken, as the genetic stocks used in the earlier studies are not available. The genetic constitutions of the three parents and their F1s are proposed. These accessions should be useful for conducting allelic tests for determining flower color loci in chickpea and for comparative studies with field pea. The seeds of these genetic stocks are maintained at the Genetic Resources and Enhancement Program at ICRISAT and are available for research purposes on request.     

The role of genes influencing the corolla in pollination of Antirrhinum majus

Studies of pollination ecology have been hindered by an absence of biochemical information about the basis of polymorphism. Using model plants and mutant lines described by molecular genetics may circumvent this difficulty. Mutation of genes controlling petal colour and petal epidermal cell shape in Antirrhinum majus was previously shown to influence fruit set. White flowers set less fruit than magenta flowers and mutants with flat petal epidermal cells set less fruit than flowers with conical cells. Here we analyse the causal pathway underlying this phenomenon through a study of floral characteristics and bee behaviour. Results indicate that bees recognized plants with magenta conical‐celled flowers at a distance and did not approach white flowers or magenta flat‐celled flowers so frequently. Petal cell shape interacted with colour in determining whether an approaching bee landed on a flower within a plot and whether a bee landing on a flower would probe it. The intrafloral temperature of flowers with conical petal cells was shown to increase with solar irradiance, unlike the intrafloral temperature of flowers with flat petal cells. The difference in fruit set may reflect pollinator discrimination between genotypes as a consequence of the effect of intrafloral temperature on nectar quality and quantity.     

The never ripe mutation blocks ethylene perception in tomato.

Seedlings of tomato fruit ripening mutants were screened for their ability to respond to ethylene. Ethylene induced the triple response in etiolated hypocotyls of all tomato ripening mutants tested except for one, Never ripe (Nr). Our results indicated that the lack of ripening in this mutant is caused by ethylene insensitivity. Segregation analysis indicated that Nr-associated ethylene insensitivity is a single codominant trait and is pleiotropic, blocking senescence and abscission of flowers and the epinastic response of petioles. In normal tomato flowers, petal abscission and senescence occur 4 to 5 days after the flower opens and precede fruit expansion. If fertilization does not occur, pedicel abscission occurs 5 to 8 days after petal senescence. If unfertilized, Nr flowers remained attached to the plant indefinitely, and petals remained viable and turgid more than four times longer than their normal counterparts. Fruit development in Nr plants was not preceded by petal senescence; petals and anthers remained attached until they were physically displaced by the expanding ovary. Analysis of engineered 1-aminocyclopropane-1-carboxylate (ACC) synthase-overexpressing plants indicated that they are phenotypic opposites of Nr plants. Constitutive expression of ACC synthase in tomato plants resulted in high rates of ethylene production by many tissues of the plant and induced petiole epinasty and premature senescence and abscission of flowers, usually before anthesis. There were no obvious effects on senescence in leaves of ACC synthase overexpressers, suggesting that although ethylene may be important, it is not sufficient to cause tomato leaf senescence; other signals are clearly involved.     

大豆花荚败育期间的植物激素变化

大豆开花结荚期,不同发育阶段的幼蕾与花荚的脱落率不同,其中以花后５ｄ内的幼荚脱落最严重。与败育花荚相比,正常花荚中的干物质积累量均较高。细胞分裂素（ＤＨＺＲｓ,ＺＲｓ,ｉＰＡ）含量也较高,花后３～５ｄ的幼荚中表现更明显。脱落酸（ＡＢＡ）则是以败育幼蕾及花后３～５ｄ的幼荚中含量较高。不同发育阶段的大豆生殖器官中,正常开放花中的玉米赤霉烯酮（ＺＥＮ）含量最高     

Penetrance and expressivity of the gene for double podding in chickpea

The double-pod per peduncle trait is known to contribute to increased seed yield in chickpea (Cicer arietinum L.). A cross was made between the single-podded variety ICCV 2 and the double-podded variety JG 62 in 1993. Penetrance and expressivity of the gene for double podding was studied in an F2 population and F10 recombinant inbred lines (RILs) of this cross. Homozygous recessive allele of this gene (ss) governs the production of double flowers and pods per peduncle. Results indicated that the s allele has unstable penetrance and variable expressivity. The penetrance of this allele was 53% for the F2 and 84.5% for the RILs. The ranges for the expression of this trait among the penetrant F2 individuals and the penetrant RILs were 1.1-14.8% and 0.1-33.0%. These were 8.3-30.8% for early sown and 17.1-68.7% for the late sown double-podded parent JG 62. Thus it appears that the allele shows greater penetrance and enhanced expressivity under soil moisture stress. In the F2 the seed yield advantage of the double-podded over the single-podded plants was 18%, whereas among the RILs it was 7%. The increased number of pods and seeds contributed to the higher yield. However, there was a slight decrease in seed size of the double-podded genotypes. An increase in the size of seed may have a role in the decreased penetrance and expressivity of this allele among the double-podded segregants of the ICCV 2 x JG 62 chickpea cross.     

SUPERMAN, a regulator of floral homeotic genes in Arabidopsis.

We describe a locus, SUPERMAN, mutations in which result in extra stamens developing at the expense of the central carpels in the Arabidopsis thaliana flower. The development of superman flowers, from initial primordium to mature flower, is described by scanning electron microscopy. The development of doubly and triply mutant strains, constructed with superman alleles and previously identified homeotic mutations that cause alterations in floral organ identity, is also described. Essentially additive phenotypes are observed in superman agamous and superman apetala2 double mutants. The epistatic relationships observed between either apetala3 or pistillata and superman alleles suggest that the SUPERMAN gene product could be a regulator of these floral homeotic genes. To test this, the expression patterns of AGAMOUS and APETALA3 were examined in superman flowers. In wild-type flowers, APETALA3 expression is restricted to the second and third whorls where it is required for the specification of petals and stamens. In contrast, in superman flowers, APETALA3 expression expands to include most of the cells that would normally constitute the fourth whorl. This ectopic APETALA3 expression is proposed to be one of the causes of the development of the extra stamens in superman flowers. The spatial pattern of AGAMOUS expression remains unaltered in superman flowers as compared to wild-type flowers. Taken together these data indicate that one of the functions of the wild-type SUPERMAN gene product is to negatively regulate APETALA3 in the fourth whorl of the flower. In addition, superman mutants exhibit a loss of determinacy of the floral meristem, an effect that appears to be mediated by the APETALA3 and PISTILLATA gene products.     

FLD interacts with CO to affect both flowering time and floral initiation in Arabidopsis thaliana.

fld and co, both with significantly delayed flowering, are characterized as late-flowering mutations in Arabidopsis thaliana. Double mutants between fld-2 and co-3 were generated and the phenotypes characterized. Double mutants flower later than both single mutant parents, suggesting that there is an additive effect. In addition, the formation of flowers in double mutants was altered and showed a novel phenotype. Double mutant flowers contained a much longer stalk (pedicel). Sepals and petals were absent. Several leaf-like structures were produced in the position normally occupied by sepals and the organ numbers were reduced. The carpels were morphologically normal. The stamens produced were usually aborted in the early stage, thus, the flowers were sterile. The additive phenotype observed in double mutants provides evidence to support that these two genes, FLD and CO, are not only involved in rosette-to-inflorescence transition but also involved in the flower formation. This result also indicates that FLD and CO promote the reproductive program through two different pathways.     

The Delayed Terminal Flower Phenotype Is Caused by a Conditional Mutation in the CENTRORADIALIS Gene of Snapdragon

The snapdragon (Antirrhinum majus) centroradialis mutant (cen) is characterized by the development of a terminal flower, thereby replacing the normally open inflorescence by a closed inflorescence. In contrast to its Arabidopsis counterpart, terminal flower1, the cen-null mutant displays an almost constant number of lateral flowers below the terminal flower. Some partial revertants of an X-radiation-induced cen mutant showed a delayed formation of the terminal flower, resulting in a variable number of lateral flowers. The number of lateral flowers formed was shown to be environmentally controlled, with the fewer flowers formed under the stronger flower-inducing conditions. Plants displaying this "Delayed terminal flower" phenotype were found to be heterozygous for a mutant allele carrying a transposon in the coding region and an allele from which the transposon excised, leaving behind a 3-bp duplication as footprint. As a consequence, an iso-leucine is inserted between Asp148 and Gly149 in the CENTRORADIALIS protein. It is proposed that this mutation results in a low level of functional CEN activity, generating a phenotype that is more similar to the Arabidopsis Terminal flower phenotype.     

Isolation of Temperature-Sensitive Mutants of Arabidopsis thaliana That Are Defective in the Redifferentiation of Shoots

Three temperature-sensitive mutants of Arabidopsis thaliana that were defective in the redifferentiation of shoots were isolated as tools for the study of organogenesis. M3 lines were constructed by harvesting M3 seeds separately from each M2 plant. Comparative examination of shoot redifferentiation in root explants of 2700 M3 lines at 22[deg]C (permissive temperature) and at 27[deg]C (restrictive temperature) led to the identification of seven temperature-sensitive mutant lines. Genetic tests of three of the seven mutant lines indicated that temperature-sensitive redifferentiation of shoots in these three lines resulted from single, nuclear, recessive mutations in three different genes, designated SRD1, SRD2, and SRD3. The morphology of root explants of srd mutants cultured at the restrictive temperature suggests that the products of these SRD genes function at different stages of the redifferentiation of shoots.     

Activation tagging,a novel tool to dissect the functions of a gene family

In a screen for morphological mutants from the T1 generation of approximately 50 000 activation-tagging lines, we isolated four dominant mutants that showed hyponastic leaves, downward-pointing flowers and decreased apical dominance. We designated them isoginchaku (iso). The iso-1D and iso-2D are allelic mutants caused by activation of the AS2 gene. The T-DNAs were inserted in the 3' downstream region of AS2. Iso-3D and iso-4D are the other allelic mutants caused by activation of the ASL1/LBD36 gene. These two genes belong to the AS2 family that is composed of 42 genes in Arabidopsis. The only recessive mutation isolated from this gene family was of AS2, which resulted in a leaf morphology mutant. Applying reverse genetics using a database of activation-tagged T-DNA flanking sequences, we found a dominant mutant that we designated peacock1-D (pck1-D) in which the ASL5/LBD12 gene was activated by a T-DNA. The pck1-D mutants have lost apical dominance, have epinastic leaves and are sterile. These results strongly suggest that activation tagging is a powerful mutant-mining tool especially for genes that make up a gene family.     

Restriction Fragment Length Polymorphism and Random Amplified Polymorphic DNA Analysis of Chickpea Accessions

Genetic diversity analysis was carried out in chickpea accessions using restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) techniques. RFLP analysis using 26 Pst I sub-genomic clones on ten chickpea accessions in 130 probe-enzyme combinations detected polymorphism with only two clones. Pst I clones, CG 141 detected polymorphism in ICC 4918 and Pusa 209 while CG 500 detected polymorphism in Pusa 261, ILC 26 and in ILC 13326. These clones detected very few polymorphic markers. Analysis using 10 Eco RI clones on twelve chickpea accessions have shown better hybridisation signal and one clone detected polymorphism in Pusa 256. RFLP analysis of both cultivated and wild Cicer species using heterologous DNA probe Cab3C revealed polymorphism only in wild Cicer species (Cicer reticulatum L., JM 2100). RAPD analysis of 13 chickpea accessions which includes mutants of C 235 and E100Y showed greater degree of polymorphism with 1 - 5 unique DNA bands for all the accessions. Phylogenetic analysis of the RAPD data helped to group the accessions. C 235 and its mutants were found to be closely grouped while E100Y and its mutant E100Ym grouped apart. Desi and kabuli chickpea accessions however, could not be separately grouped. This revised version was published online in July 2006 with corrections to the Cover Date.     

One for all and all for one: retention of colour‐unchanged old flowers increases pollinator attraction in a hermaphroditic plant

• Long‐lived flowers increase pollen transfer rates, but these entail high water and carbon maintenance costs. The retention of pollinated and reward‐free old flowers enhances pollinator visitation to young receptive flowers by increasing floral display size. This mechanism is associated with acropetal inflorescences or changes in flower colour and openness, but the retention of unchanging solitary flowers remains overlooked.
• We examined pollination‐dependent variation in floral longevity and determined stigmatic receptivity, pollen viability and pollen removal rates among flower ages in Kielmeyera regalis, a Neotropical savanna shrub. We also evaluated the effects of floral display size on pollinator visitation rates. Lastly, we determined whether old flowers are unvisited and exclusively increase pollinator attraction to young flowers through flower removal experiments.
• Regardless of pollination treatment, flowers lasted fully open with no detectable physical changes for 3 days. Over time, stigmas remained receptive but >95% of pollen was removed. Pollinator visitation significantly increased with floral display size and intermediate percentages (15–30%) of newly opened flowers. Accordingly, the retention of reward‐free and unvisited old flowers increased young flower–pollinator interaction.
• Our results reveal the importance of a prolonged floral longevity in increasing pollinator attraction toward newly opened receptive flowers without changes in flower colour and form. We conclude that the retention of pollinated, reward‐free and unvisited colour‐unchanged old flowers in K. regalis is a strategy that counteracts the water use costs associated with the maintenance of large flowers with increased mate opportunities in a pollen‐limited scenario.