Pigeonpea breeding in eastern and southern Africa: challenges and opportunities

Pigeonpea (Cajanus cajan [L.] Millspaugh) is an important multipurpose grain legume crop primarily grown in tropical and subtropical areas of Asia, Africa and Latin America. In Africa, the crop is grown for several purposes including food security, income generation, livestock feed and in agroforestry. Production in Eastern and Southern Africa (ESA) is however faced with many challenges including limited use of high‐yielding cultivars, diseases and pests, drought, under‐investment in research and lack of scientific expertise. The aim of this review is to highlight the challenges facing pigeonpea breeding research in ESA and the existing opportunities for improving the overall pigeonpea subsector in the region. We discuss the potential of the recently available pigeonpea genomic resources for accelerated molecular breeding, the prospects for conventional breeding and commercial hybrid pigeonpea, and the relevant seed policies, among others, which are viewed as opportunities to enhance pigeonpea productivity.     

沙棘籽油对牛的皮肤伤口愈合疗效的研究

本研究旨在通过比较沙棘籽油（Hippophae rhamnoides）和防腐软膏（5％碘伏）、草本软膏（印度草药研究与供应有限公司）及一种刺激性温和的基础药膏（液体石蜡）,来研究沙棘籽油对牛的皮肤伤口愈合效果。这个实验是将12头雄性牛犊分为4组,6头牛都有皮肤伤口,3头胸背部脊椎区的两边都用当地的镇痛药。伤口分别用液体石蜡、5％的碘伏软膏、沙棘籽油（沙棘油）及草本软膏处理28d,并分为Ⅰ、Ⅱ、Ⅲ、Ⅳ四组。不同处理的功效是建立在对经过0d、3d、7d、10d、14d和28d的不同临床血液参数的研究基础之上的。在研究中,所有组的心脏、呼吸率和血液学的各种参数及肛温都维持在正常的生理极限内。沙棘油和草本软膏对伤口处理的临床研究中发现,在伤口愈合的初期,伤口保持了一定的干燥度,并且抑制了炎症初期伤口的恶化。不同的组之间,在持续28d的实验中,用草本软膏和沙棘油做处理的组中伤口的愈合率最高。这些组的伤口试验同时还表现出早治疗,早治愈的特征。但是,在草本软膏和沙棘油处理组中,草本软膏组处理的伤口的整体愈合能力还是略优于沙棘油的疗效。     

QTLs for chlorophyll and chlorophyll fluorescence parameters in barley under post-flowering drought

Drought is one of the major factors limiting barley yields in many developing countries worldwide. The identification of molecular markers linked to genes controlling drought tolerance in barley is one way to improve breeding efficiency. In this study, we analyzed the quantitative trait loci (QTL) controlling chlorophyll content and chlorophyll fluorescence in 194 recombinant inbred lines (RILs) developed from the cross between the cultivar ‘Arta’ and Hordeum spontaneum 41-1. Five traits, chlorophyll content, and four chlorophyll fluorescence parameters, namely initial fluorescence (F_o), maximum fluorescence (F_m), variable fluorescence (F_v), and maximum quantum efficiency of PSII (F_v/F_m) which are related to the activity of the photosynthetic apparatus, were measured under well-watered and drought stress conditions at post-flowering stage. QTL analysis identified a total of nine and five genomic regions, under well-watered and drought stress conditions, respectively, that were significantly associated with the expression of the five target traits at post-flowering stage. No common QTL was detected except one for chlorophyll content, which was identified in both growth conditions, demonstrating that the genetic control of the expression of the traits related to photosynthesis differed under different water conditions. A QTL for F_v/F_m, which is related to the drought tolerance of photosynthesis was identified on chromosome 2H at 116 cM in the linkage map under drought stress. This QTL alone explained more than 15% of phenotypic variance of maximum quantum yield of PSII, and was also associated with the expression of four other traits. In addition, another QTL for F_v/F_m was also located on the same chromosome (2H) but at 135.7 cM explaining around 9% of the phenotypic variance under drought conditions. The result presented here suggest that two major loci, located on chromosome 2H, are involved in the development of functional chloroplast at post-flowering stage for drought tolerance of photosynthesis in barley under drought stress. If validated in other populations, chlorophyll fluorescence parameters could be used as selection criteria for drought tolerance.     

The drivers and methodologies for exploiting wild <Emphasis Type="Italic">Cajanus</Emphasis> genome in pigeonpea breeding

With the exception of Cajanus cajan (L.) Millspaugh (pigeonpea), the remaining species of genus Cajanus have not been domesticated. For millennia these taxa have persisted in natural habitats through self-sown seeds. These wild species are an asset for sustaining future pigeonpea breeding programmes since they contain certain traits (genes) that are necessary for encountering various breeding challenges related to crop improvement and adaptation. In this review we identify the key traits from wild Cajanus species, and discuss various physical and genetic constraints encountered in their utilization in introgression breeding. Some noteworthy achievements recorded from inter-specific breeding programmes in pigeonpea are also discussed. These include the development of (1) high protein (>?28%) genotypes (2) cytoplasmic nuclear male sterility systems (3) highly (>?95%) self-pollinating genotypes, and (4) resistance sources to sterility mosaic disease, nematodes, salinity, photo-insensitivity, pod borers, podfly, bruchids, and Phytophthora blight. To help pigeonpea breeders engaged in inter-specific breeding programmes, we suggest the division of the secondary gene pool germplasm into two sub-group/tiers on the basis of ease in hybridization.     

Ovulation,Pregnancy Rate and Early Embryonic Development in Vernal Transitional Mares Treated with Equine‐ or Porcine‐FSH

The objective of this study was to compare the efficacy of purified equine‐ and porcine‐FSH treatment regimes in mares in early vernal transition. Mares (n = 22) kept under ambient light were examined ultrasonographically per‐rectum, starting January 30th. They were assigned to one of two treatment groups using a sequential alternating treatment design when a follicle ≥ 25 mm was detected. In the eFSH group, mares were treated twice daily with equine‐FSH, and in the pFSH group mares were treated twice daily with porcine‐FSH; treatments were continued until follicle(s) ≥ 35 mm, and 24 h later hCG was administered. Oestrous mares were inseminated with fresh semen and examined for pregnancy on days 11–20 post‐ovulation. In the eFSH group, 11/11 (100%) mares developed follicle(s) ≥ 35 mm, 8/11 (73%) ovulated and 6/8 (75%) conceived. In the pFSH group, 5/11 (45%) developed follicle(s) ≥ 35 mm, 4/11 (36%) ovulated and 3/4 (75%) conceived. Treatment with eFSH resulted in a greater ovarian stimulation; higher number of pre‐ovulatory‐sized follicles, higher number of ovulations and higher number of embryos (p < 0.05). Following ovulation, serum progesterone concentrations were correlated with the number of CLs and supported early embryonic development; maternal recognition of pregnancy occurred in all pregnant mares. We concluded that eFSH can be used to effectively induce follicular growth and ovulation in vernal transitional mares; however, if bred, diagnosis and management of twins’ pregnancies would be required prior to day 16 because of the increased risk of multiple embryos per pregnancy. Conversely, the current pFSH treatment regime cannot be recommended.     

Effect of Roscovitine‐Treated Donor Cells on Development of Porcine Cloned Embryos

Synchronization of the donor cell cycle is an important factor for successful animal cloning by nuclear transfer. To improve the efficiency of porcine cloning, in the present report, we evaluated effects of contact inhibition, serum starvation and roscovitine treatment of donor cells on in vitro and in vivo developmental potency of cloned porcine embryos. Fibroblasts derived from a porcine foetus at day 30 of gestation were isolated and cultured to 70% confluency. Then, cells were either cultured to 100% confluency for contact inhibition, or cultured in 0.5% serum for 72 h for serum starvation or with 15 μm roscovitine for 24 h. Cells were most effectively synchronized at G0/G1 in the serum starvation group (87.5%) compared with the contact inhibition and roscovitine treatment groups (76.3% and 79.9% respectively p < 0.05). However, after somatic cell nuclear transfer followed by in vitro culture, the serum starvation group showed a significantly lower blastocyst formation rate (5.6%) compared with the contact inhibition and roscovitine treatment groups (11.6% and 20.0% respectively). Differential expression of apoptosis‐related genes and the level of apoptosis in each treatment group explain the variation in developmental competence among the groups. Significantly higher level of apoptosis was observed in the serum starvation group. On the other hand, the roscovitine treatment group shows the lowest level of apoptosis and the best in vitro development among the groups. Cloned embryos derived from roscovitine‐treated donor cells were transferred to surrogate pigs. Three healthy live piglets were produced. In conclusion, we suggest that roscovitine treatment of donor cells improves development of cloned porcine embryos and can raise the efficiency of cloned piglet production.     

A comparative assessment of the utility of PCR-based marker systems in pearl millet

A set of 22 pearl millet inbred lines including the parents of eleven mapping populations, was screened with 627 markers including 100 pearl millet genomic SSRs (gSSRs), 60 pearl millet EST-SSRs (eSSRs), 410 intron sequence haplotypes (ISHs), and 57 exon sequence haplotypes (ESHs). In all, 267 (59%) of the markers were informative for at least one of the 11 mapping populations, which segregate for traits like drought and salinity tolerance; host plant resistance to downy mildew, rust and blast; fertility restoration and sterility and maintenance of cytoplasmic male sterility etc. An average of 116 polymorphic markers was identified per mapping population. The average PIC values and number of profiles (P) per polymorphic marker were: gSSRs (PIC = 0.62, P = 6.1), ISHs (PIC = 0.39, P = 2.6), eSSRs (PIC = 0.36, P = 3.1) and ESHs (PIC = 0.35, P = 3.1). A high correlation (r > 0.97, P < 0.05) was observed between the patterns of diversity exposed by the different marker systems. The polymorphic markers identified are suitable for the de novo construction, or the supplementation of pearl millet linkage maps. The genetic relationships identified among the panel of inbred lines may be useful in designing strategies to improve the use of available genetic variation in the context of pearl millet breeding.     

Sensitivity of boreal forest carbon balance to soil thaw

We used eddy covariance; gas-exchange chambers; radiocarbon analysis; wood, moss, and soil inventories; and laboratory incubations to measure the carbon balance of a 120-year-old black spruce forest in Manitoba, Canada. The site lost 0.3 +/- 0.5 metric ton of carbon per hectare per year (ton C ha-1 year-1) from 1994 to 1997, with a gain of 0.6 +/- 0.2 ton C ha-1 year-1 in moss and wood offset by a loss of 0.8 +/- 0.5 ton C ha-1 year-1 from the soil. The soil remained frozen most of the year, and the decomposition of organic matter in the soil increased 10-fold upon thawing. The stability of the soil carbon pool ( approximately 150 tons C ha-1) appears sensitive to the depth and duration of thaw, and climatic changes that promote thaw are likely to cause a net efflux of carbon dioxide from the site.     

Use of immature seed germination and single seed descent for rapid genetic gains in pigeonpea

Long duration required for generation advancement in pigeonpea [Cajanus cajan (L.) Millsp] is one of the major bottlenecks in realizing rapid genetic gains. Therefore, a technology for rapid generation turnover is warranted to facilitate the development of new cultivars and recombinant inbred lines. Breeding of early‐maturing cultivars has now opened up the possibility of rapid generation advance (RGA) in this crop. This paper reports the development of an RGA technology that integrates the germination of immature seeds with single seed descent method of breeding. The results showed that immature 35‐day‐old seeds can be used successfully to turn over a generation of pigeonpea with 100% seed germination. These way 3/4 successive generations can be grown within a year. The methodology presented in this study will accelerate the breeding process for breeding cultivars and develop rapidly the materials required for genomics research in pigeonpea.     

Maturation Status, Protein Synthesis and Developmental Competence of Oocytes Derived from Lambs and Ewes

The efficacy of oocyte selection for in vitro embryo production depends on the abundance and diameter of follicles, cumulus layers around the oocytes and subsequent fertilization. Application of `ovum pick-up' technique allows us to utilize partially matured oocytes for embryo production even from juvenile subjects. To compare their developmental competence, oocytes derived from lambs and ewes and cultured in maturation medium for up to 26 h were assessed at 2 h intervals by confocal microscopy after chromatin and microtubulin-specific fluorochrome labelling. Lamb oocytes reached second meiotic metaphase (MII) at lower numbers at 24 h (60.0%) and 26 h (28.6%) whereas 85.7% of adult-derived oocytes attained MII status by 24 h of maturation. Radiolabelling of oocyte proteins revealed higher incorporation of [³⁵S-]-methionine and [³⁵S]-cysteine in adult-derived oocytes compared to lamb oocytes. Although the cleavage rate of lamb oocytes was similar to that of ewe oocytes, the proportion reaching blastocyst stage was significantly lower (p < 0.05) in the lamb-derived oocytes. However, blastocysts from both types of oocytes displayed similar cell lineage allocations to inner cell mass and trophectoderm.