Water and Nutrient Use Efficiency in Diploid, Tetraploid and Hexaploid Wheats

Three diploid (Triticum boeoticum, AA; Aegilops speltoides, BB and Ae. tauschii, DD), two tetraplold (T. dlcoccoides,AABB and T. dicoccon, AABB) and one hexaploid (T. vulgare, AABBDD) varieties of wheat, which are very important in the evolution of wheat were chosen in this study. A pot experiment was carried out on the wheat under different water and nutrient conditions (i) to understand the differences in biomass, yield, water use efficiency (WUE), and nutrient (N, P and K) use efficiency (uptake and utilization efficiency) among ploldles in the evolution of wheat; (ii) to clarify the effect of water and nutrient conditions on water and nutrient use efficiency; and (iii) to assess the relationship of water and nutrient use efficiency in the evolution of wheat. Our results showed that from diploid to tetraploid then to hexaploid during the evolution of wheat, both root biomass and above-ground biomass increased initially and then decreased. Water consumption for transpiration decreased remarkably, correlating with the decline of the growth period, while grain yield, harvest index, WUE, N, P and K uptake efficiency, and N, P and K utilization efficiency increased significantly. Grain yield, harvest index and WUE decreased in the same order: T.vulgare ＞ T. dicoccon ＞ T. dicoccoides ＞ Ae. tauschii ＞ Ae. speltoides ＞ T. boeoticum. Water stress significantly decreased root biomass, above-ground biomass, yield, and water consumption for transpiration by 47-52%, butremarkably increased WUE. Increasing the nutrient supply increased wheat above-ground biomass, grain yield,harvest index, water consumption for transpiration and WUE under different water levels, but reduced root biomass under drought conditions. Generally, water stress and low nutrient supply resulted in the lower nutrientuptake efficiency of wheat. However, water and nutrient application had no significant effects on nutrient utilization efficiency, suggesting that wheat nutrient utilization efficiency is mainly controlled by genotypes. Compared to theother two diploid wheats, Ae. squarrosa (DD) had significant higher WUE and nutrient utilization efficiency, Indicating that the D genome may carry genes controlling high efficient utilization of water and nutrient. Significant relationships were found between WUE and N, P and K utilization efficiency.     

Factors influencing shoot regeneration from cotyledons of tetraploid <Emphasis Type="Italic">Isatis indigotica</Emphasis> fort

Summary An efficient in vitro plant regeneration system from cotyledons was established in tetraploid Isatis indigotica Fort. Factors influencing shoot regeneration from cotyledons, including culture medium type, combinations of plant growth regulators, and sucrose concentrations in the medium, as well as illumination were investigated. Murashige and Skoog's (MS) medium was found to be best for promoting shoot regeneration, followed by Gamborg's B₅ and White's medium. The highest shoot regeneration frequency was achieved from cotyledons cultured on MS medium supplemented with 2.0 mgl⁻¹ (8.9 μM) 6-benzyladenine and 1.0 mgl⁻¹ (5.4 μM) α-naphthaleneacetic acid (NAA), with 97.9% regeneration, associated with a high number of multiple shoots developed per explant (8.6 shoots per explant). A sucrose concentration of 3% present in the medium and light conditions were beneficial for shoot regeneration. The shoots developed were rooted in a half-strength MS medium supplemented with 1.0 mgl⁻¹ (5.4 μM) NAA and successfully transplanted in soil in pots with over 85% survival. The establishment of an efficient plant regeneration procedure from cotyledons provides a basis for the rapid in vitro multiplication of tetraploid Isatis indigotica Fort., one of the most extensively used medicinal plants in China currently under great shortage.     

Cytological studies on the genus Holcoglossum (Orchidaceae)

The karyotypes of ten species of Holcoglossum (Orchidaceae), a highly endangered and diversified genus from China, were investigated to study the infrageneric relationships, biogeography, and speciation patterns in the Hengduan Mountains. The karyotype formulae of the studied species are as follows: 2 n  = 38 = 20m + 18sm in H. subulifolium , 2 n  = 38 = 22m + 16sm in H. amesianum , 2 n  = 38 = 26m + 12sm (6 SAT) in H. lingulatum , 2 n  = 38 = 26m + 12sm in H. wangii , 2 n  = 38 = 10m + 28sm in H. kimballianum , 2 n  = 38 = 14m + 22sm + 2st in H. flavescens , 2 n  = 38 = 24m + 12sm + 2st in H. rupestre , 2 n  = 38 = 14m + 20sm + 4st in H. sinicum , 2 n  = 38 = 16m + 14sm + 8st in H. weixiense , and 2 n  = 76 in H. tsii . The karyotypes of two tropical species, H. amesianum and H. subulifolium , are the most primitive in the genus, whereas those of four temperate alpine species, H. sinicum , H. rupestre , H. weixiense , and H. flavescens , are more advanced. H. tsii is a tetraploid and H. rupestr e may be one of its ancestors. The low frequency of polyploidy in Holcoglossum in the Hengduan Mountains region supports the conclusion that chromosome stasis during rapid speciation is common there.  © 2007 The Linnean Society of London. Botanical Journal of the Linnean Society , 2007, 154 , 283–288.     

六个石蒜居群的核型及四倍体石蒜的发现

染色体与核型的变化是植物系统发育和进化的一个重要方面。石蒜属Lycoris植物特别是石蒜L.radiata在染色体数目和核型上存在较大的变异。通过对不同居群的石蒜核型研究,可以为石蒜和石蒜属植物的核型演化及演化机制提供一些重要的基础资料。本文对分布于中国安徽省和浙江省的6个石蒜居群进行了细胞学研究。结果表明,6个石蒜居群的染色体数目和核型分别为：霍山居群2n=44=28st＋8t＋8T,2n=22=6st＋12t＋4T;黄山居群2n=22=22t,2n=22=18st＋4t,2n=21=12st＋7t＋2T;滁州居群2n=33=33t;马鞍山居群2n=33=18st＋15T,2n=25=1m＋20st＋2t＋2T;宣城居群2n=22=20st＋2T,2n=21=1m＋20st;杭州居群2n=22=12st＋4t＋6T,2n=21=18st＋3t。其中,部分居群的核型类型为首次报道;并首次发现了四倍体的石蒜居群。此外,对石蒜的核型进化和多倍体起源进行了初步探讨。     

Simple and efficient production of mice derived from embryonic stem cells aggregated with tetraploid embryos

Six newly derived hybrid mouse embryonic stem (ES) cell lines and two inbred ES cell lines were tested for their ability to produce completely ES cell-derived mice by aggregation of ES cells with tetraploid embryos. Forty-five ES cell-tetraploid pups were generated from six hybrid ES cell lines and no pups from two inbred ES cell lines. These pups were found to have increased embryonic and placental weights than control mice. Twenty-two pups survived to adulthood and produced normal offsprings, and the other 23 pups died of several reasons including respiratory distress, abdomen ulcer-like symptoms, and foster failure. The 22 adult ES cell-tetraploid mice were completely ES cell-derived as judged by coat color and germline transmission, only two of them was found to have tetraploid component in liver, blood, and lung as analyzed by microsatellite loci. Our data suggested that genetic heterozygosity is a crucial factor for postnatal survival of ES cell-tetraploid mice, and tetraploid embryo aggregation using hybrid ES cells is a simple and efficient procedure for immediate generation of targeted mouse mutants from genetically modified ES cell clones, in contrast to the standard protocol, which involves the production of chimeras and several breeding steps.     

Comparative histology of the testes of the spined loach Cobitis taenia L. and natural allotetraploids of Cobitis (Pisces, Cobitidae)

The spined loach Cobitis taenia L. creates exclusively diploid and mixed diploid–polyploid populations. Allotriploid females, which co-exist with C. taenia or C. elongatoides and a few tetraploid males and females dominate in most Cobitis mixed populations. They reproduce gynogenetically and produce triploid eggs that are stimulated to development by sperm from Cobitis males. Some of these eggs are fertilized, which leads to the production of bisexual tetraploids. Males of C. taenia (2n = 48) from a diploid population in Lake Klawój, Northern Poland (46 individuals) and from a mixed Cobitis population in the Bug River, Eastern Poland (7 individuals), and three tetraploid males (4n = 98) from the same mixed population were examined. All the fish were analyzed karyologically and histologically. Tubules with cysts of the testes of C. taenia from both populations were filled with germ cells at various developmental stages. Among fishes from Lake Klawój sperm maturation in batches simultaneous with the batch spawning of C. taenia females was found. The testes of the loach C. taenia, from a mixed population in the Bug River, were filled with spermatozoa over the entire reproductive season. Sperm maturation in batches was not observed. Sperm maturation in batches seems to be only connected with a few diploid males in this population. So, a continuous process of spermatogenesis in their testes is required. Only in the testes of all tetraploid Cobitis males were cells characteristic of the early stages of spermatogenesis observed, i.e. without spermatids and spermatozoa. Furthermore, the histological sections of the testis of a male captured in August, revealed fragments with connective tissue between the germ cells. However the participation of tetraploid, infertile Cobitis males in the process of reproduction in the investigated mixed population remains controversial. The results obtained so far, reveal that even the infertile sperm of tetraploid males may induce gynogenesis in Cobitis triploid females.     

Tetraploid cells from cytokinesis failure induce aneuploidy and spontaneous transformation of mouse ovarian surface epithelial cells

Most ovarian cancers originate from the ovarian surface epithelium and are characterized by aneuploid karyotypes. Aneuploidy, a consequence of chromosome instability, is an early event during the development of ovarian cancers. However, how aneuploid cells are evolved from normal diploid cells in ovarian cancers remains unknown. In the present study, cytogenetic analyses of a mouse syngeneic ovarian cancer model revealed that diploid mouse ovarian surface epithelial cells (MOSECs) experienced an intermediate tetraploid cell stage, before evolving to aneuploid (mainly near-tetraploid) cells. Using long-term live-cell imaging followed by fluorescence in situ hybridization (FISH), we demonstrated that tetraploid cells originally arose from cytokinesis failure of bipolar mitosis in diploid cells, and gave rise to aneuploid cells through chromosome mis-segregation during both bipolar and multipolar mitoses. Injection of the late passage aneuploid MOSECs resulted in tumor formation in C57BL/6 mice. Therefore, we reveal a pathway for the evolution of diploid to aneuploid MOSECs and elucidate a mechanism for the development of near-tetraploid ovarian cancer cells.     

Chk1 suppresses bypass of mitosis and tetraploidization in p53-deficient cancer cells

Many cancer cells are unable to maintain a numerically stable chromosome complement. It is well established that aberrant cell division can generate progeny with increased ploidy, but the genetic factors required for maintenance of diploidy are not well understood. Using an isogenic model system derived by gene targeting, we examined the role of Chk1 in p53-proficient and -deficient cancer cells. Targeted inactivation of a single CHK1 allele in stably diploid cells caused an elevated frequency of mitotic bypass if p53 was naturally mutated or experimentally disrupted by homologous recombination. CHK1-haploinsufficient, p53-deficient cells frequently underwent sequential rounds of DNA synthesis without an intervening mitosis. These aberrant cell cycles resulted in whole-genome endoreduplication and tetraploidization. The unscheduled bypass of mitosis could be suppressed by targeted reversion of a p53 mutation or by exogenous expression of Cdk1. In contrast, the number of tetraploid cells was not increased in isogenic cell populations that harbor hypomorphic ATR mutations, suggesting that suppression of unscheduled mitotic bypass is a distinct function of Chk1. These results are consistent with a recently described role for Chk1 in promoting the expression of genes that promote cell cycle transitions and demonstrate how Chk1 might prevent tetraploidization during the cancer cell cycle.     

A review of ploidy in the genus Prosopis (Leguminosae)

The genus Prosopis contains 44 species of trees and shrubs, the majority of which originate in the Americas. Most species are reported to be diploid, with a somatic chromosome number of 2 n  = 28. There are rare reports of polyploidy, although it is thought that these may represent polysomaty in root tissues. However, flow cytometry has recently indicated that P. juliflora is entirely tetraploid with a somatic chromosome number of 2 n  = 56. In order to clarify the situation, an extensive review of ploidy in Prosopis was undertaken, the first of its kind. The ploidy levels of 124 samples of Prosopis from 21 countries, including both the natural and introduced ranges, were analysed using flow cytometry. In addition, a comprehensive literature review was carried out, examining 305 published ploidy values and covering 32 of the 44 species of Prosopis . Flow cytometry analysis suggests that P. juliflora is the only tetraploid species, with a somatic chromosome number of 2 n  = 4 x  = 56, whilst the remainder of the species analysed are diploid with 2 n  = 2 x  = 28, including the first report for P. articulata (2 n  = 28). A critical review of published ploidy values shows that all species of Prosopis are reported to be entirely diploid, except P. glandulosa , P. juliflora , and P. koelziana , for which both diploid and tetraploid values have been recorded. © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society , 2008, 156 , 425–438.     

四倍体刺槐的抗盐性

 以四倍体刺槐(Robinia pseudoacacia)为主要试验材料,以二倍体刺槐为对照材料,在两种盐胁迫下,对苗木的形态、生理生化、光合特性和解剖结构等指标的变化规律进行了研究。利用NaCl和Na₂SO₄盐溶液对两种刺槐进行盐处理,在30 d后每7 d处理1次,共处理4次,并在处理前和处理后每7 d进行各项指标的测定。结果发现：1)在盐胁迫下二倍体刺槐的植株生长受到强烈抑制,叶片含水量和叶绿素含量显著低于对照,有明显的盐害症状;对四倍体刺槐植株的生长影响较小,叶片含水量和叶绿素含量也与对照差异不显著,无盐害症状。2)四倍体刺槐经过盐处理后相对电导率和脯氨酸(Proline, Pro)含量虽然也稍有上升,但与对照相比未达到显著水平,而二倍体刺槐显著高于对照;同时作为保护酶系统的过氧化物酶(Peroxidase, POD)、超氧化物歧化酶(Superoxide dismutase, SOD)、过氧化氢酶(Catalase, CAT)在四倍体刺槐经盐胁迫后期也保持了较高的活性,从而提高了其抗盐性,而对盐敏感的二倍体刺槐3个保护酶活性均较低。 3)经盐胁迫后对四倍体刺槐光合特性影响不大,净光合速率（Net photosynthetic rate, P_n）和胞间CO₂浓度（Intercellular CO₂ concentration, C_i）均无显著变化,而二倍体刺槐P_n和C_i显著下降。4)经盐胁迫后四倍体刺槐在解剖结构上做出了积极的反应：叶肉栅栏组织拉长、排列更为紧密、 海绵组织变小、排列紧密。而二倍体刺槐出现了相反的现象。综合以上分析认为：四倍体刺槐具有较强的抗盐性。