Intracellular ice and cell survival in cryo-exposed embryonic axes of recalcitrant seeds of Acer saccharinum: an ultrastructural study of factors affecting cell and ice structures

Background and Aims

Cryopreservation is the only long-term conservation strategy available for germplasm of recalcitrant-seeded species. Efforts to cryopreserve this form of germplasm are hampered by potentially lethal intracellular freezing events; thus, it is important to understand the relationships among cryo-exposure techniques, water content, structure and survival.

Methods

Undried embryonic axes of Acer saccharinum and those rapidly dried to two different water contents were cooled at three rates and re-warmed at two rates. Ultrastructural observations were carried out on radicle and shoot tips prepared by freeze-fracture and freeze-substitution to assess immediate (i.e. pre-thaw) responses to cooling treatments. Survival of axes was assessed in vitro.

Key Results

Intracellular ice formation was not necessarily lethal. Embryo cells survived when crystal diameter was between 0·2 and 0·4 µm and fewer than 20 crystals were distributed per μm² in the cytoplasm. Ice was not uniformly distributed within the cells. In fully hydrated axes cooled at an intermediate rate, the interiors of many organelles were apparently ice-free; this may have prevented the disruption of vital intracellular machinery. Intracytoplasmic ice formation did not apparently impact the integrity of the plasmalemma. The maximum number of ice crystals was far greater in shoot apices, which were more sensitive than radicles to cryo-exposure.

Conclusions

The findings challenge the accepted paradigm that intracellular ice formation is always lethal, as the results show that cells can survive intracellular ice if crystals are small and localized in the cytoplasm. Further understanding of the interactions among water content, cooling rate, cell structure and ice structure is required to optimize cryopreservation treatments without undue reliance on empirical approaches.     

Homoiohydrous (recalcitrant) seeds: Developmental status,desiccation sensitivity and the state of water in axes of Landolphia kirkii Dyer

The desiccation sensitivity in relation to the stage of development was investigated in embryonic axes from the homoiohydrous (recalcitrant) seeds of Landolphia kirkii. Electrolyte leakage, used to assess membrane damage after flash (very rapid) drying, indicated that axes from immature (non-germinable) seeds were the most desiccation-tolerant, followed by those from mature seeds, while axes from seeds germinated for increasing times were progressively more desiccation-sensitive. Differential scanning calorimetry was used to study the relationship between desiccation sensitivity and the properties of water in the tissues. Axes from immature seeds had a lower content of non-freezable water than that of any other developmental stage and a higher enthalpy of melting of freezable water. For mature and immature axes electrolyte leakage increased at the point of loss of freezable water. At other developmental stages the water content at which electrolyte leakage increased markedly correlated with the other properties of the water, such as the change in the shape of the melting endotherm and the onset temperature. Ultrastructural studies of axes at the various developmental stages showed changes in the degree and pattern of vacuolation, the presence and quantities of lipid and starch, and the degree of endomembrane development. The results are discussed in relation to current hypotheses on the basis of desiccation tolerance.Abbreviation DSC differential scanning calorimetry     

Increases in the longevity of desiccation-phase developing rice seeds: response to high-temperature drying depends on harvest moisture content

Background and Aims Previous studies have suggested that the drying conditions routinely used by genebanks may not be optimal for subsequent seed longevity. The aim of this study was to compare the effect of hot-air drying and low-temperature drying on subsequent seed longevity for 20 diverse rice accessions and to consider how factors related to seed production history might influence the results.Methods Seeds of rice, Oryza sativa, were produced according to normal regeneration procedures at IRRI. They were harvested at different times [harvest date and days after anthesis (DAA), once for each accession] and dried either in a drying room (DR; 15 % relative humidity, 15 °C) or in a flat-bed heated-air batch dryer (BD; 45 °C, 8 h d^–1) for up to six daily cycles followed by drying in the DR. Relative longevity was assessed by storage at 10·9 % moisture content and 45 °C.Key Results Initial drying in the BD resulted in significantly greater longevity compared with the DR for 14 accessions (seed lots): the period of time for viability to fall to 50 % for seeds dried in the BD as a percentage of that for seeds dried throughout in the DR varied between 1.3 and 372·2 % for these accessions. The seed lots that responded the most were those that were harvested earlier in the season and at higher moisture content. Drying in the BD did not reduce subsequent longevity compared with DR drying for any of the remaining accessions.Conclusions Seeds harvested at a moisture content where, according to the moisture desorption isotherm, they could still be metabolically active (>16·2 %) may be in the first stage of the post-mass maturity, desiccation phase of seed development and thus able to increase longevity in response to hot-air drying. The genebank standards regarding seed drying for rice and, perhaps, for other tropical species should therefore be reconsidered.     

A review of the genus Raveniola (Araneae, Nemesiidae) in China, with notes on allied genera and description of four new species from Yunnan

The Chinese representatives of Raveniola Zonstein, 1987 are currently recognized to comprise seven species. Four new species - Raveniola montanasp. n. (♂♀), Raveniola shangrilasp. n. (♂), Raveniola songisp. n. (♂) and Raveniola yunnanensissp. n. (♂) - are described from the highlands of Yunnan Province, China. According to some characters (shape of the palpus, palpal tibia and tibia I in males) they can be placed together with Raveniola hebeinica Zhu, Zhang & Zhang, 1999 and with Raveniola guangxi (Raven & Schwendinger, 1995), comb. n., transferred here from Sinopesa Raven & Schwendinger, 1995. The current generic position of Raveniola xizangensis (Hu & Li, 1987) is confirmed. Other Chinese nemesiids referred previously to Raveniola are transferred to Sinopesa: Sinopesa chinensis (Kulczyński, 1901), comb. n., Sinopesa sinensis (Zhu & Mao, 1983), comb. n. and Sinopesa chengbuensis (Xu & Yun, 2002), comb. n. The relationships between these Asian genera and their relations to Afrotropical nemesiids are discussed.     

New species of springtails in the Proisotoma genus complex from Vermont and New York, USA with descriptive notes on Ballistura alpa Christiansen & Bellinger 1980 (Hexapoda, Collembola, Isotomidae)

Three new species of Isotomidae springtails are described from the Lake Champlain Basin (Vermont and New York, USA), Lake Willoughby and Greater Averril Pond in Vermont. Subisotoma joyceisp. n. and Scutisotoma champisp. n. were collected in sandy beaches whereas Ballistura rossisp. n. was found only in a constructed wetland built and managed by the University of Vermont. Scutisotoma champisp. n. was found in Lakes Champlain and Willoughby, and Greater Averril Pond and is probably present in most lakes and large ponds in the area. Subisotoma joyceisp. n. was found only along the southern and eastern coast of South Hero, and the mainland coast facing eastern South Hero. Ballistura alpa is redescribed and transferred to the genus Pachyotoma based on the absence of tibiotarsal seta B4/B5, the presence of secondary cuticular granules, 4 prelabral setae, a full complement of guard setae on labial papilla E and in having a bifurcate outer maxillary lobe with 4 sublobal setae.     

One new species of the subgenus Hexatoma (Eriocera) Macquart (Diptera,Limoniidae) from China with a key to Chinese species

One new species of the subgenus Eriocera Macquart, 1838, Hexatoma (Eriocera) cleopatroides Men, sp. n. (Southern China: Anhui) is described and illustrated. A key to all of 78 known species from China in the subgenus is provided, which was solely based on literatures. The new species is similar to Hexatoma (Eriocera) cleopatra Alexander, 1933, but distinguishes from the latter by the prescutum entirely black with two ill-defined gray stripes, by the legs with fore and middle femora brown in basal half, black in apical half, with hind femora brown in basal one-fourth, and by the wings with cells c and sc more yellowish brown than the ground color.     

A new species of the brown lacewing genus Zachobiella Banks from China (Neuroptera,Hemerobiidae) with a key to species

The genus Zachobiella Banks, 1920 is reviewed and a new species Zachobiella yunanica sp. n. described from China. All species found in China are redescribed, and Zachobiella submarginata Esben-Petersen, 1929 is recorded from China for the first time. A key to the adults of Zachobiella is provided.     

New species of subgenus Tipula (Sivatipula) from China,with redescription of T. (S.) parvauricula and a key to all known species of the Oriental Region (Diptera,Tipulidae, Tipula)

Species of Tipula (Sivatipula) biprocessa sp. n. from Guangxi, China is described and illustrated as new in the subgenus Tipula (Sivatipula) Alexander, 1964. Tipula (Sivatipula) parvauricula Alexander, 1941 is redescribed and illustrated based on additional morphological characters. Semen pump of this subgenus is discussed. A key to all described species in this group is compiled.     

Zinc induces distinct changes in the metabolism of reactive oxygen and nitrogen species (ROS and RNS) in the roots of two Brassica species with different sensitivity to zinc stress

Background and Aims Zinc (Zn) is an essential micronutrient naturally present in soils, but anthropogenic activities can lead to accumulation in the environment and resulting damage to plants. Heavy metals such as Zn can induce oxidative stress and the generation of reactive oxygen and nitrogen species (ROS and RNS), which can reduce growth and yield in crop plants. This study assesses the interplay of these two families of molecules in order to evaluate the responses in roots of two Brassica species under high concentrations of Zn.Methods Nine-day-old hydroponically grown Brassica juncea (Indian mustard) and B. napus (oilseed rape) seedlings were treated with ZnSO₄ (0, 50, 150 and 300 µm) for 7 d. Stress intensity was assessed through analyses of cell wall damage and cell viability. Biochemical and cellular techniques were used to measure key components of the metabolism of ROS and RNS including lipid peroxidation, enzymatic antioxidants, protein nitration and content of superoxide radical (O2·−), nitric oxide (NO) and peroxynitrite (ONOO⁻).Key Results Analysis of morphological root damage and alterations of microelement homeostasis indicate that B. juncea is more tolerant to Zn stress than B. napus. ROS and RNS parameters suggest that the oxidative components are predominant compared with the nitrosative components in the root system of both species.Conclusions The results indicate a clear relationship between ROS and RNS metabolism as a mechanism of response against stress caused by an excess of Zn. The oxidative stress components seem to be more dominant than the elements of the nitrosative stress in the root system of these two Brassica species.     

The Tetramorium squaminode species group (Hymenoptera,Formicidae) in the Arabian Peninsula,with a new record from the Kingdom of Saudi Arabia and keys to Arabian species

The Arabian species of the Tetramorium squaminode-group are treated. Tetramorium squaminode Santschi, 1911 is recorded for the first time from the Kingdom of Saudi Arabia and the Arabian Peninsula. Keys to the two Arabian species of the Tetramorium squaminode-group, Tetramorium latinode Collingwood & Agosti, 1996 and Tetramorium squaminode, based on worker and queen castes, are given and a regional distribution map is provided. Notes on habitats of Tetramorium squaminode are presented.     

Aenictus hoelldobleri sp. n., a new species of the Aenictus ceylonicus group (Hymenoptera,Formicidae) from China,with a key to the Chinese members of the group

Aenictus is the most species-rich genus of army ants in the subfamily Dorylinae and one of the most species rich ant genera in China and the world. In this paper, a new species of the Aenictus ceylonicus group, Aenictus hoelldobleri sp. n., is described and illustrated based on the worker caste. The new species occurs in the subtropical forests of south-east China and is morphologically most similar to Aenictus henanensis Li & Wang, 2005 and Aenictus wudangshanensis Wang, 2006. Aenictus hoelldobleri sp. n. can be distinguished from both species by the shape of the subpetiolar process. The new species also resembles Aenictus lifuiae Terayama 1984 and Aenictus thailandianus Terayama & Kubota, 1993 but clearly differs in various features of the cuticular sculpture. A key to the Chinese species of the Aenictus ceylonicus group based on the worker caste is provided, which may help to reassess and clarify the taxonomic status of the abundant Chinese records of the true Aenictus ceylonicus (Mayr, 1866), a species which almost certainly does not occur in China. Several new locality records are given, among them the first record of Aenictus watanasiti Jaitrong & Yamane, 2013 from China.