Research progress of plant cytogenetics in Jiangsu province.

Cytogenetics was established based on the "Chromosome theory of inheritance", proposed by Boveri and Sutton and evidenced by Morgan's lab in early stage of the 20 th centrary. With rapid development of related research areas, especially molecular genetics, cytogenetics developed from traditional into a new era, molecular cytogenetics in late 1960s. Featured by an established technique named DNA in situ hybridization (ISH), molecular cytogenetics has been applied in various research areas. ISH provids vivid and straightforward figures showing the virtual presence of DNA, RNA or proteins. In combination with genomics and cell biology tools, ISH and derived techniques have been widely used in studies of the origin, evolution, domestication of human, animal and plant, as well as wide hybridization and chromosome engineering. The physical location and order of DNA sequences revealed by ISH enables the detection of chromosomal re-arrangments among related species and gaps of assembled genome sequences. In addition, ISH using RNA or protein probes can reveal the location and quantification of transcripted RNA or translated protein. Since the 1970s, scientists from universities or institutes belonging to the Jiangsu Society of Genetics have initiated cytogenetics researches using various plant species. In recent years, research platforms for molecular cytogenetics have also been well established in Nanjing Agricultural University, Yangzhou University, Nanjing Forestry University, Jiangsu Xuhuai Academy of Agricultural Sciences, and Jiangsu Normal University. The application of molecular cytogenetics in plant evolution, wide hybridization, chromosome engineering, chromosome biology, genomics has been successful. Significant progresses have been achieved, both in basic and applied researches. In this paper, we will review main research progresses of plant cytogenetics in Jiangsu province, and discuss the potential development of this research area.

Transcriptomic and proteomic analysis reveals mechanisms of low pollen-pistil compatibility during water lily cross breeding.

BACKGROUND: In water lily (Nymphaea) hybrid breeding, breeders often encounter non-viable seeds, which make it difficult to transfer desired or targeted genes of different Nymphaea germplasm. We found that pre-fertilization barriers were the main factor in the failure of the hybridization of Nymphaea. The mechanism of low compatibility between the pollen and stigma remains unclear; therefore, we studied the differences of stigma transcripts and proteomes at 0, 2, and 6 h after pollination (HAP). Moreover, some regulatory genes and functional proteins that may cause low pollen-pistil compatibility in Nymphaea were identified. RESULTS: RNA-seq was performed for three comparisons (2 vs 0 HAP, 6 vs 2 HAP, 6 vs 0 HAP), and the number of differentially expressed genes (DEGs) was 8789 (4680 were up-regulated), 6401 (3020 were up-regulated), and 11,284 (6148 were up-regulated), respectively. Using label-free analysis, 75 (2 vs 0 HAP) proteins (43 increased and 32 decreased), nine (6 vs 2 HAP) proteins (three increased and six decreased), and 90 (6 vs 0 HAP) proteins (52 increased and 38 decreased) were defined as differentially expressed proteins (DEPs). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses revealed that the DEGs and DEPs were mainly involved in cell wall organization or biogenesis, S-adenosylmethionine (SAM) metabolism, hydrogen peroxide decomposition and metabolism, reactive oxygen species (ROS) metabolism, secondary metabolism, secondary metabolite biosynthesis, and phenylpropanoid biosynthesis. CONCLUSIONS: Our transcriptomic and proteomic analysis highlighted specific genes, incuding those in ROS metabolism, biosynthesis of flavonoids, SAM metabolism, cell wall organization or biogenesis and phenylpropanoid biosynthesis that warrant further study in investigations of the pollen-stigma interaction of water lily. This study strengthens our understanding of the mechanism of low pollen-pistil compatibility in Nymphaea at the molecular level, and provides a theoretical basis for overcoming the pre-fertilization barriers in Nymphaea in the future.

Factors affecting quantity of pollen dispersal of spray cut chrysanthemum (Chrysanthemum morifolium).

BACKGROUND: Spray cut chrysanthemum is a vital flower with high ornamental value and popularity in the world. However, the excessive quantity of pollen dispersal of most spray cut chrysanthemum is an adverse factor during its flowering stage, and can significantly reduce its ornamental value and quickly shorten its vase life. More seriously, excessive pollen grains in the air are usually harmful to people, especially for those with pollen allergies. Therefore, in order to obtain some valuable information for developing spray cut chrysanthemum with less-dispersed or non-dispersed pollen in the future breeding programs, we here investigated the factors affecting quantity of pollen dispersal of spray cut chrysanthemum with four cultivars, i.e. 'Qx-097', 'Noa', 'Qx-115', and 'Kingfisher', that have different quantity of pollen dispersal. RESULTS: 'Qx-097' with high quantity of pollen dispersal has 819 pollen grains per anther, 196.4 disk florets per inflorescence and over 800,000 pollen grains per inflorescence. The corresponding data for 'Noa' with low quantity of pollen dispersal are 406, 175.4 and over 350,000, respectively; and 219, 144.2 and nearly 160,000 for 'Qx-115' without pollen dispersal, respectively. 'Kingfisher' without pollen dispersal has 202.8 disk florets per inflorescence, but its anther has no pollen grains. In addition, 'Qx-097' has a very high degree of anther cracking that nearly causes a complete dispersal of pollen grains from its anthers. 'Noa' has a moderate degree of anther cracking, and pollen grains in its anthers are not completely dispersed. However, the anthers of 'Qx-115' and 'Kingfisher' do not crack at all. Furthermore, microsporogenesis and pollen development are normal in 'Qx-097', whereas many microspores or pollen degenerate in 'Noa', most of them abort in 'Qx-115', and all of them degrade in 'Kingfisher'. CONCLUSIONS: These results suggest that quantity of pollen dispersal in spray cut chrysanthemum are mainly determined by pollen quantity per anther, and capacity of pollen dispersal. Abnormality during microsporogenesis and pollen development significantly affects pollen quantity per anther. Capacity of pollen dispersal is closely related to the degree of anther dehiscence. The entire degeneration of microspore or pollen, or the complete failure of anther dehiscence can cause the complete failure of pollen dispersal.

Identification of chrysanthemum (Chrysanthemum morifolium) self-incompatibility.

There has been a heated argument over self-incompatibilityof chrysanthemum (Chrysanthemum morifolium) among chrysanthemum breeders. In order to solve the argument, we investigated pistil receptivity, seed set, and compatible index of 24 chrysanthemum cultivars. It was found that the 24 cultivars averagely had 3.7-36.3 pollen grains germinating on stigmas at 24 hours after self-pollination through the fluorescence microscope using aniline blue staining method. However, only 10 of them produced self-pollinated seeds, and their seed sets and compatible indexes were 0.03-56.50% and 0.04-87.50, respectively. The cultivar "Q10-33-1" had the highest seed set (56.50%) and compatible index (87.50), but ten of its progeny had a wide range of separation in seed set (0-37.23%) and compatible index (0-68.65). The results indicated that most of chrysanthemum cultivars were self-incompatible, while a small proportion of cultivars were self-compatible. In addition, there is a comprehensive separation of self-incompatibility among progeny from the same self-pollinated self-compatible chrysanthemum cultivar. Therefore, it is better to emasculate inflorescences during chrysanthemum hybridization breeding when no information concerning its self-incompatibility characteristics is available. However, if it is self-incompatible and propagated by vegetative methods, it is unnecessary to carry out emasculation when it is used as a female plant during hybridization breeding.

Morphological, physiological, and structural responses of two species of artemisia to NaCl stress.

Effects of salt stress on Artemisia scoparia and A. vulgaris "Variegate" were examined. A. scoparia leaves became withered under NaCl treatment, whereas A. vulgaris "Variegate" leaves were not remarkably affected. Chlorophyll content decreased in both species, with a higher reduction in A. scoparia. Contents of proline, MDA, soluble carbohydrate, and Na(+) increased in both species under salt stress, but A. vulgaris "Variegate" had higher level of proline and soluble carbohydrate and lower level of MDA and Na(+). The ratios of K(+)/Na(+), Ca(2+)/Na(+), and Mg(2+)/Na(+) in A. vulgaris "Variegate" under NaCl stress were higher. Moreover, A. vulgaris "Variegate" had higher transport selectivity of K(+)/Na(+) from root to stem, stem to middle mature leaves, and upper newly developed leaves than A. scoparia under NaCl stress. A. vulgaris "Variegate" chloroplast maintained its morphological integrity under NaCl stress, whereas A. scoparia chloroplast lost integrity. The results indicated that A. scoparia is more sensitive to salt stress than A. vulgaris "Variegate." Salt tolerance is mainly related to the ability of regulating osmotic pressure through the accumulation of soluble carbohydrates and proline, and the gradient distribution of K(+) between roots and leaves was also contributed to osmotic pressure adjustment and improvement of plant salt tolerance.

Factors influencing fecundity in experimental crosses of water lotus (Nelumbo nucifera Gaertn.) cultivars.

BACKGROUND: Breeding programs for the water lotus (Nelumbo nucifera) are hampered by an inability to account for variation in seed set associated with crosses between different cultivars. We studied seed set in two reciprocal crosses between lotus cultivars ('Guili' × 'Aijiangnan' and 'Molingqiuse' × 'Qinhuaiyanzhi') to obtain insights into factors that govern fecundity in these experimental hybrids. Pollen viability, stigma receptivity and embryo development were compared for each hybrid and reciprocal cross. RESULTS: Pollen viability of the individual cultivars ranged from 4.1% to 20.2%, with the highest level (>11.9%) for all cultivars observed from the earliest collected grains (05:00-06:00 a.m.). Stigmatic pollen germination peaked at 4 h after pollination and varied from 4.8 to 60.6 grains per stigma among the crosses. Production of normal embryos ranged from 7.6% to 58.8% at 1 d after pollination and from 0 to 25% by 11 d after pollination. Seed set in crosses (0.2-23.3%) was generally lower than in open-pollinated plants (8.4-26.5%). Similar to the germination results, seed set was substantially reduced in both reciprocal crosses. CONCLUSIONS: These results suggested that poor pollen fertility, low stigma receptivity, and embryo abortion were responsible for the failure of the crosses 'Molingqiuse' × 'Qinhuaiyanzhi', 'Qinhuaiyanzhi' × 'Molingqiuse', and 'Aijiangnan' × 'Guili'.

Pollen viability, pistil receptivity, and embryo development in hybridization of Nelumbo nucifera Gaertn.

Seed set is usually low and differs for different crosses of flower lotus (Nelumbo nucifera Gaertn.). The reasons remain unknown, and this has a negative impact on lotus breeding. To determine the causes, we carried out two crosses of flower lotus, that is, "Jinsenianhua" × "Qinhuaihuadeng" and "Qinhuaihuadeng" × "Jinsenianhua" and pollen viability, pistil receptivity, and embryo development were investigated. The pollen grains collected at 05:00-06:00 hrs had the highest viability, and the viabilities of "Jinsenianhua" and "Qinhuaihuadeng" were 20.6 and 15.7%, respectively. At 4 h after artificial pollination, the number of pollen grains germinating on each stigma reached a peak: 63.0 and 17.2 per stigma in "Jinsenianhua" × "Qinhuaihuadeng" and "Qinhuaihuadeng" × "Jinsenianhua", respectively. At 1 d after artificial pollination, the percentages of normal embryos in the two crosses were 55.0 and 21.9%, respectively; however, at 11 d after pollination, the corresponding percentages were 20.8 and 11.2%. Seed sets of the two crosses were 17.9 and 8.0%, respectively. The results suggested that low pistil receptivity and embryo abortion caused low seed set in "Qinhuaihuadeng" × "Jinsenianhua", whereas low fecundity of "Jinsenianhua" × "Qinhuaihuadeng" was mainly attributable to embryo abortion.

Effects of CO(2) enrichment on growth and development of Impatiens hawkeri.

The effects of CO(2) enrichment on growth and development of Impatiens hawkeri, an important greenhouse flower, were investigated for the purpose of providing scientific basis for CO(2) enrichment to this species in greenhouse. The plants were grown in CO(2)-controlled growth chambers with 380 (the control) and 760 (CO(2) enrichment) µmol · mol(-1), respectively. The changes in morphology, physiology, biochemistry, and leaf ultrastructure of Impatiens were examined. Results showed that CO(2) enrichment increased flower number and relative leaf area compared with the control. In addition, CO(2) enrichment significantly enhanced photosynthetic rate, contents of soluble sugars and starch, activities of peroxidase (POD), superoxide dismutase (SOD), and ascorbate peroxidase (APX), but reduced chlorophyll content and malondialdehyde (MDA) content. Furthermore, significant changes in chloroplast ultrastructure were observed at CO(2) enrichment: an increased number of starch grains with an expanded size, and an increased ratio of stroma thylakoid to grana thylakoid. These results suggest that CO(2) enrichment had positive effects on Impatiens, that is, it can improve the visual value, promote growth and development, and enhance antioxidant capacity.

[Changes of growth and yield characters of five varieties (lines) of tea-applied chrysanthemum under different planting dates and pinching patterns]. / 不同定植期和摘心方案下5个品种(系)茶用菊生长和产量性状的变化.

To identify the high yield planting date and pinching pattern, we compared the yields of five new varieties (lines) of tea-applied chrysanthemum, the ‘Suju 10’, ‘Suju 12’, ‘Suju 13’, ‘CH1-44’ and ‘CH5-13’. A field experiment with split-split plot design was carried out. The main plot treatments including three (early, middle and late) planting dates, split-plot treatments including 5 new varieties (lines), and split-split-plot treatments including four different kinds of pinching schemes. The results showed that the yield of ‘CH5-13’ and ‘Suju 13’ was relatively higher among the five varieties (lines), followed by ‘CH1-44’ and ‘Suju10’, and ‘Suju 12’ showed the lowest yield. Among the five varieties (lines), planting date on May 27th and two-time pinching treatments showed the highest values of plant height, crown width, flower number, flower diameter, fresh mass, yield per plant and yield per unit area. Compared with the planting date on May 7th and June 13th, the above indexes increased by 16.0% and 19.0%, 18.0% and 22.8%, 36.7% and 42.2%, 11.1% and 2.3%, 13.0% and 4.0%, 47.8% and 36.6%, 48.5% and 36.7%, respectively. With the pinching time postponed, plant height decreased. Compared with the no pinching treatment, plant height of the two-time pinching treatment decreased by 50.2%, and values of crown width, flowers number, the flower fresh mass, the yield per plant and the yield per unit area were highest, with increases of 17.0%, 29.1%, 5.5%, 34.0% and 34.8%, respectively. The impact performance of three main factors on the growth and yield of the tea-applied chrysanthemum was in order of planting date>varieties>pinching time.

[Effects of crude toxin from Alternaria alternata on the growth and physiological metabolism of chrysanthemum 'Jinba' seedlings]. / é“¾æ ¼å­¢èŒç²—æ¯’ç´ å¯¹èŠèŠ±'神马'幼苗生长及生理代谢的影响.

The chrysanthemum black spot caused by Alternaria alternata significantly reduced the quality and yield of chrysanthemum. The crude toxin secreted by A. alternata in the metabolic process have elopathic effects on plants, which is the main pathogenic factor for the occurrence of chrysanthemum black spot. The pathogenic fungi A. alternate was isolated from chrysanthemum black spot leaves, The effects of crude toxin on plant height, stem diameter, root length, resistant material content, membrane relative permeability, polyphenol oxidase (PPO), peroxidase (POD), phenylalanine ammonia-lyase (PAL) in different treatments of chrysanthemum 'Jinba' seedlings were investigated. The results showed that the crude toxin of A. alternata had an inhibition effect on plant height, stem diameter and root length. The toxin concentration was positively correlated with the inhibitory effect. 14 days after crude toxin treatment, plant height, stem diameter and root length were significantly inhibited, with an reduction of 28.9%, 21.4% and 23.3%, respectively. The cell membrane permeability of leaf increased with the toxin concentration. Under the same toxin concentration, the cell membrane permeability first increased and then decreased with the treatment duration. The contents of soluble protein, malondialdehyde (MDA) and proline in leaves were significantly increased after treatment with the toxin solution. The increases of PAL, POD and PPO were the most significant in 10 times A. alternatacrude toxin treatment. The pathogenicity of A. alternate crude toxin to the chrysanthemum 'Jinba' seedlings was mainly through inhibi-ting the normal growth of roots and stems, destructing the root cell membrane permeability and increasing the contents of MDA, normal soluble sugar and proline, and promoting the activities of PAL, POD, PPO in leaf tissues.

[Effects of bio-organic fertilizer and fungicide application on continuous cropping obstacles of cut chrysanthemum].

Abstract: Fusarium wilt is a soil borne disease caused by plant continuous cropping in monoculture Chrysanthemum morifolium 'Youxiang' monoculture not only declines plant quality and yield but also decreases soil enzymes and soil microbial diversity over successive cultivation. In this article, the effects of fungicide (Carbendazim MBC), antifungal enhanced bio-organic fertilizer (BOF), and their combined application on the quality and soil enzymes activities of Chrysanthemum morifolium 'Youxiang' in continuous cropping systems were investigated. The results showed that both bioorganic fertilizer (BOF) and fungicide (MBC) single application could effectively prevent the occurrence of Fusarium wilt disease of cut chrysanthemum. Bio-organic fertilizer application was more effective on root activity, soil enzymes activities and quality (shoot height, stem diameter, leaf SPAD value, ray floret number, shoot fresh mass) improvement of cut chrysanthemum, while fungicide single application was responsible for soil enzymatic activities suppression to some extent. The combined application treatment (MBC+BOF) showed the best effects on quality improvement and soil enzyme activities promotion.

[Effects of stem numbers per ground area on the quality of standard cut Chrysanthemum morifolium in greenhouse: simulation model].

In order to understand the effects of stem numbers per ground area on the quality of standard cut Chrysanthemum morifolium, an experiment with different cultivars, different stem numbers per plant, different planting densities, and different planting dates was conducted in a greenhouse in Shanghai in 2005 and 2006. The effects of stem numbers per ground area on the canopy leaf area index and external quality of standard cut C. morifolium were quantified using the experimental data. Based on the physiological product of thermal effectiveness and PAR (PETP) the canopy absorbed, a model for predicting the effects of stem numbers per ground area on the quality of standard cut C. morifolium was developed, and validated with independent experimental data. The results showed that with the increase of stem numbers per ground area, the leaf area index increased, whereas plant height, stem diameter, leaf number, and flower diameter decreased. The model gave satisfactory predictions of the quality of standard cut C. morifolium cultivated with different stem numbers and planting density. The coefficient of determination (R2) and relative prediction error (RSE) based on the 1:1 line for fresh mass per stem, plant height, stem diameter, leaf number, flower diameter, and the number of qualified stem harvested per ground area were 0.95, 0.96, 0.94, 0.91, 0.81 and 0.97, and 16.1%, 10.1%, 12.8%, 13.4%, 15.9%, 16.1% , respectively. The model developed in this study could be used for the optimization of light and temperature management for standard cut C. morifolium cultivated with different stem numbers and planting densities in greenhouse.

[Effects of transplanting date and density on appearance quality of greenhouse single-flower cut chrysanthemum].

Transplanting date and density are the important factors affecting the appearance quality of chrysanthemum. The study on the greenhouse single-flower cut chrysanthemum (Chrysanthemum morifolium cv. Shenma) showed that within the ranges of test transplanting date and density, the plant height and neck length increased, while the leaf number per plant, stem diameter, plant fresh mass and flower diameter decreased with the delay of transplanting date and the increase of transplanting density. No effect of transplanting density was observed on plant height. For the production of single-flower cut chrysanthemum in non-heated greenhouse in Shanghai, the optimal transplanting date and density to achieve the top rank of quality (rank A) were the middle ten days of August and 64 plants x m(-2), and those to achieve the second rank of quality (rank B) were from mid August to early September and 72-80 plants x m(-2), respectively. The results obtained in this study offered references in establishing the prediction model of greenhouse single-flower cut chrysanthemum appearance quality based on light, temperature, and transplanting date and density.

[Quality prediction model of greenhouse standard cut chrysanthemum based on light-temperature effect].

Based on the effects of light and temperature on chrysanthemum quality and the experiments with different chrysanthemum varieties and planting dates, a quality prediction model of greenhouse standard cut chrysanthemum with the physiological product of thermal effectiveness and PAR (PTEP) as the measurement scale was developed and validated. The results showed that the predicted results, including the number of unfolding leaves, leaf area, plant height, stem diameter, internode length and flower diameter, accorded well with the observed ones, and the determination coefficient (R2) and relative prediction error (RSE) based on 1 : 1 line were 0.99, 0.98, 0.98, 0.92, 0.87 and 0.88, and 5.5%, 6.5%, 5.9%, 4.1%, 11.2%, 12.4%, respectively. This model was of high precision and practicable, which could be used in optimizing the light and temperature management for greenhouse standard cut chrysanthemum production.