Sham S. Goyal

Arbuscular mycorrhizas, microbial communities, nutrient availability, and soil aggregates in organic tomato production

Effects of arbuscular mycorrhzal (AM) fungi on plant growth and nutrition are well-known, but their effects on the wider soil biota are less clear. This is in part due to difficulties with establishing appropriate non-mycorrhizal controls in the field. Here we present results of a field experiment using a new approach to overcome this problem. A previously well-characterized mycorrhizal defective tomato mutant (rmc) and its mycorrhizal wildtype progenitor (76R MYC+) were grown at an organic fresh market tomato farm (Yolo County, CA). At the time of planting, root in-growth cores amended with different levels of N and P, were installed between experimental plants to study localized effects of mycorrhizal and non-mycorrhizal tomato roots on soil ecology. Whilst fruit yield and vegetative production of the two genotypes were very similar at harvest, there were large positive effects of colonization of roots by AM fungi on plant nutrient contents, especially P and Zn. The presence of roots colonized by AM fungi also resulted in improved aggregate stability by increasing the fraction of small macroaggregates, but only when N was added. Effects on the wider soil community including nematodes, fungal biomass as indicated by ergosterol, microbial biomass C, and phospholipid fatty acid (PLFA) profiles were less pronounced. Taken together, these data show that AM fungi provide important ecosystem functions in terms of plant nutrition and aggregate stability, but that a change in this one functional group had only a small effect on the wider soil biota. This indicates a high degree of stability in soil communities of this organic farm.

Enhancement of the salt tolerance of Triticum turgidum L. by the Kna1 locus transferred from the Triticum aestivum L. chromosome 4D by homoeologous recombination

Durum wheat, Triticum turgidum L. (2n= 4x=28, genome formula AABB) is inferior to bread wheat, T. aestivum L. (2n=6x=42, genome formula AABBDD), in the ability to exclude Na+ under salt strees, in the ratio of the accumulated K+ to Na+ in the leaves under salt stress, and in tolerance of salt stress. Previous work showed that chromosome 4D has a major effect on Na+ and K+ accumulation in the leaves of bread wheat. The 4D chromosome was recombined with chromosome 4B in the genetic background of durum wheat. The recombinants showed that Na+ exclusion and enhanced K+/Na+ ratio in the shoots were controlled by a single locus, Kna1, in the long arm of chromosome 4D. The recombinant families were grown in the field under non-saline conditions and two levels of salinity to determine whether Kna1 confers salt tolerance. Under salt stress, the Kna1 families had higher K+/Na+ ratios in the flag leaves and higher yields of grain and biomass than the Kna1- families and the parental cultivars. Kna1 is, therefore, one of the factors responsible for the higher salt tolerance of bread wheat relative to durum wheat. The present work provides conceptual evidence that tolerance of salt stress can be transferred between species in the tribe Triticeae.

Molecular characterization of durum and common wheat recombinant lines carrying leaf rust resistance (Lr19) and yellow pigment (Y) genes from Lophopyrum ponticum

Chromosome 7E from Lophopyrum ponticum carries a valuable leaf rust resistant gene designated Lr19. This gene has not been widely used in common wheat breeding because of linkage with the yellow pigment gene Y. This gene tints flour yellow, reducing its appeal in bread making. However, a high level of yellow pigment is desirable in durum wheat breeding. We produced 97 recombinant chromosomes between L. ponticum transfer 7D.7E#1 and its wheat homoeologues, using the ph1b mutation that promotes homoeologous pairing. We characterized a subset of 37 of these lines with 11 molecular markers and evaluated their resistance to leaf rust and the abundance of yellow pigment. The Lr19 gene was mapped between loci Xwg420 and Xmwg2062, whereas Y was mapped distal to Xpsr687, the most distal marker on the long arm of chromosome 7. A short terminal 7EL segment translocated to 7A, including Lr19 and Y (line 1-23), has been transferred to durum wheat by backcrossing. The presence of this alien segment significantly increased the abundance of yellow pigment. The Lr19 also conferred resistance to a new durum leaf rust race from California and Mexico that is virulent on most durum wheat cultivars. The new durum lines with the recombinant 7E segment will be useful parents to increase yellow pigment and leaf rust resistance in durum wheat breeding programs. For the common wheat breeding programs, we selected the recombinant line 1-96, which has an interstitial 7E segment carrying Lr19 but not Y. This recombinant line can be used to improve leaf rust resistance without affecting flour color. The 7EL/7DL 1-96 recombinant chromosome did not show the meiotic self-elimination previously reported for a 7EL/7BL translocation.

Effect of nitrate pulses on the nitrate-uptake rate, synthesis of mRNA coding for nitrate reductase, and nitrate-reductase activity in the roots of barley seedlings

Using pulses of nitrate, instead of the permanent presence of external nitrate, to induce the nitrate-assimilating system in Hordeum vulgare L., we demonstrated that nitrate can be considered as a trigger or signal for the induction of nitrate uptake, the appearance of nitratereductase activity and the synthesis of mRNA coding for nitrate reductase. Nitrate pulses stimulated the initial rate of nitrate uptake, even after subsequent cultivation in N-free medium, and resulted in a higher acceleration of the uptake rate in the presence of nitrate than in its absence.

Engineering of interstitial foreign chromosome segments containing the K(+)/Na (+) selectivity gene Kna1 by sequential homoeologous recombination in durum wheat.

Targeted homoeologous recombination mediated by the absence of the Ph1 locus is currently the most efficient technique by which foreign genes can be introgressed into polyploid wheat species. Because intra-arm homoeologous double cross-overs are rare, introgressed foreign genes are usually on terminal foreign chromosome segments. Since the minimum length of such a segment is determined by the position of a gene in the chromosome, large chromosome segments with undesirable genetic effects are often introgressed. Introgression of foreign genes on short interstitial segments based on two cycles of homoeologous recombination is described here. The utility of the technique is demonstrated by the introgression of the Kna1 locus, which controls K+/Na+ selectivity in T. aesivum L., on short interstitial segments of chromosome 4D into chromosome 4B of Triticum turgidum L. The level of recombination in a homoeologous segment is not significantly affected by a juxtaposed proximal homologous segment in the absence of the Ph1 locus.

Isolation and characterization of a low phytic acid rice mutant reveals a mutation in the rice orthologue of maize MIK

Using a forward genetics approach, we isolated two independent low phytic acid (lpa) rice mutants, N15-186 and N15-375. Both mutants are caused by single gene, recessive non-lethal mutations, which result in approximately 75% (N15-186) and 43% (N15-375) reductions in seed phytic acid (inositol hexakisphosphate). High-performance liquid chromatography and GC–MS analysis of seed extracts from N15-186 indicated that, in addition to phytic acid, inositol monophosphate was significantly reduced whereas inorganic phosphorus and myo-inositol were greatly increased when compared with wild-type. The changes observed in N15-186 resemble those previously described for the maize lpa3 mutant. Analysis of N15-375 revealed changes similar to those observed in previously characterized rice lpa1 mutants (i.e. significant reduction in phytic acid and corresponding increase in inorganic phosphorus with little or no change in inositol phosphate intermediates or myo-inositol). Further genetic analysis of the N15-186 mutant indicated that the mutation, designated lpa N15-186, was located in a region on chromosome 3 between the microsatellite markers RM15875 and RM15907. The rice orthologue of maize lpa3, which encodes a myo-inositol kinase, is in this interval. Sequence analysis of the N15-186 allele of this orthologue (Os03g52760) revealed a single base pair change (C/G to T/A) in the first exon of the gene, which results in a nonsense mutation. Our results indicate that lpa N15-186 is a mutant allele of the rice myo-inositol kinase (OsMIK) gene. Identification and characterization of lpa mutants, such as N15-186, will facilitate studies on the regulation of phytic acid biosynthesis and accumulation and help address questions concerning the contribution of the inositol lipid-dependent and independent biosynthetic pathways to the production of seed phytic acid.

The rice OsLpa1 gene encodes a novel protein involved in phytic acid metabolism

The rice low phytic acid 1 (lpa1) mutant was originally identified using a forward genetics approach. This mutant exhibits a 45% reduction in rice seed phytic acid with a molar-equivalent increase in inorganic phosphorus; however, it does not appear to differ significantly in productivity from its wild-type progenitor. A second lpa1 mutant was identified from additional screening for high seed inorganic phosphorus phenotypes. Using a positional cloning strategy, we identified a single candidate gene at the rice Lpa1 locus. Sequence analysis of the candidate gene from the lpa1 mutants revealed two independent mutations (a single base pair substitution and a single base pair deletion) that confirmed the identification of this candidate as the rice low phytic acid 1 gene, OsLpa1. The OsLpa1 gene has three splice variants. The location and nature of the two mutations suggests that these lesions only affect the translation of the predicted protein derived from the longest transcript. The proteins encoded by OsLpa1 do not have homology to any of the inositol phosphate metabolism genes recently characterized in plants, although there is homology to 2-phosphoglycerate kinase, an enzyme found in hyperthermophilic methanogens that catalyzes the formation of 2,3-bisphosphoglycerate from 2-phosphoglycerate. OsLpa1 represents a novel gene involved in phytic acid metabolism.

Survey of aliphatic glucosinolates in Sicilian wild and cultivated Brassicaceae

In the frame of the activities carried out to exploit Sicilian local cultivars of brassicas, we focused our attention on some of the potential health compounds of various local cruciferous crops. These compounds are of interest to improve the quality of the produce with the aim to develop new cultivars capable of providing functional foods able to prevent disease. In this context, we surveyed for the presence of specific glucosinolates in local cultivars of broccoli, cauliflower, kale, and in some wild species widespread in Sicily, using as control various commercial cultivars. Glucosinolate composition varied extensively among species and crops of the same species, such as cauliflower, broccoli and kale. Cultivar variation for glucosinolate profile was also observed for some crops. For example, Sicilian cultivars of cauliflower possessing colored curds displayed a high content of glucosinolates, glucoraphanin in particular, compared to white curd commercial cultivars. Also some wild species had a high content of other glucosinolates.

Simultaneous determination of arsenite, arsenate, selenite and selenate in waters using suppressed ion chromatography with ultraviolet absorbance detection

Using Dionex AS4A anion-exchange column, with micromembrane supressor and UV detector at 195 nm, arsenite, arsenate, selenite and selenate can be determined simultaneously in water samples. The mole fraction of sodium carbonate in the eluent as well as eluent concentration affected the retention times especially for the divalent anions. An eluent of 2.5 mM Na2CO3 and 0.75 mM NaHCO3 was found to be the most suitable for sample analysis. The responses for all ions tested was linear in 0–5 mg 1−1 range. The system can tolerate up to 1600 mg 1−1 of SO2−4, before affecting arsenate peaks. Selenate peak area was not affected due to SO2−4 concentration up to 2000 mg 1−1. The detection limits using 100-μl loop were 0.1 mg 1−1 for AsO−2 and SeO2−3 and 0.25 mg 1t-1 AsO3−4 and SeO2−4.

Long-Term Reuse of Drainage Waters of Varying Salinities for Crop Irrigation in a Cotton-Safflower Rotation System in the San Joaquin Valley of California—A Nine Year Study: I. Cotton (Gossypium hirsutum L.)

Summary Use of saline drainage water for crop irrigation was evaluated as a means of decreasing its volume. Results of a nine-year crop rotation (cotton-cotton-safflower, × 3) in which only the cotton was irrigated with drainage water of 400, 1,500, 3,000, 4,500, 6,000, and 9,000 ppm total dissolved salts are presented. The different salinity levels of irrigation waters were achieved by mixing nonsaline canal water (400 ppm) and saline drainage water. Cotton lint yields were not affected by increased salinity level of the irrigation water for the first two years. Detrimental effects became evident in the third cotton crop with increasing severity in later years. In the fifth year of cotton (seventh year of the study), lint yields were adversely affected by waters of salinity greater than 3,000 ppm. However, fiber quality remained unaffected at all levels of irrigation water salinity. The reductions in lint yield appeared to be a function of time and the salinity level of applied water. Shoot height and biomass were reduced by the irrigation water salinity before lint yields. Stand establishment appeared to be the most sensitive to salinity and was perhaps the main reason for yield reduction. Increase in irrigation water salinity increased Na+ content of leaf blades and petioles and decreased K+/Na+ ratio of leaf blades and petioles. The study showed that irrigation waters of up to 3,000 ppm salinity may be used for four years without any yield reductions, as long as some leaching occurs through preplant irrigations with low salinity water. Data on crop growth and development and ionic content collected over the nine year period are presented.