Exploiting breakdown in nonhost effector-target interactions to boost host disease resistance.

Plants are resistant to most microbial species due to nonhost resistance (NHR), providing broad-spectrum and durable immunity. However, the molecular components contributing to NHR are poorly characterised. We address the question of whether failure of pathogen effectors to manipulate nonhost plants plays a critical role in NHR. RxLR (Arg-any amino acid-Leu-Arg) effectors from two oomycete pathogens, Phytophthora infestans and Hyaloperonospora arabidopsidis, enhanced pathogen infection when expressed in host plants (Nicotiana benthamiana and Arabidopsis, respectively) but the same effectors performed poorly in distantly related nonhost pathosystems. Putative target proteins in the host plant potato were identified for 64 P. infestans RxLR effectors using yeast 2-hybrid (Y2H) screens. Candidate orthologues of these target proteins in the distantly related non-host plant Arabidopsis were identified and screened using matrix Y2H for interaction with RxLR effectors from both P. infestans and H. arabidopsidis. Few P. infestans effector-target protein interactions were conserved from potato to candidate Arabidopsis target orthologues (cAtOrths). However, there was an enrichment of H. arabidopsidis RxLR effectors interacting with cAtOrths. We expressed the cAtOrth AtPUB33, which unlike its potato orthologue did not interact with P. infestans effector PiSFI3, in potato and Nicotiana benthamiana. Expression of AtPUB33 significantly reduced P. infestans colonization in both host plants. Our results provide evidence that failure of pathogen effectors to interact with and/or correctly manipulate target proteins in distantly related non-host plants contributes to NHR. Moreover, exploiting this breakdown in effector-nonhost target interaction, transferring effector target orthologues from non-host to host plants is a strategy to reduce disease.

N-Acyl-Homoserine Lactone Primes Plants for Cell Wall Reinforcement and Induces Resistance to Bacterial Pathogens via the Salicylic Acid/Oxylipin Pathway.

The ability of plants to monitor their surroundings, for instance the perception of bacteria, is of crucial importance. The perception of microorganism-derived molecules and their effector proteins is the best understood of these monitoring processes. In addition, plants perceive bacterial quorum sensing (QS) molecules used for cell-to-cell communication between bacteria. Here, we propose a mechanism for how N-acyl-homoserine lactones (AHLs), a group of QS molecules, influence host defense and fortify resistance in Arabidopsis thaliana against bacterial pathogens. N-3-oxo-tetradecanoyl-l-homoserine lactone (oxo-C14-HSL) primed plants for enhanced callose deposition, accumulation of phenolic compounds, and lignification of cell walls. Moreover, increased levels of oxylipins and salicylic acid favored closure of stomata in response to Pseudomonas syringae infection. The AHL-induced resistance seems to differ from the systemic acquired and the induced systemic resistances, providing new insight into inter-kingdom communication. Consistent with the observation that short-chain AHLs, unlike oxo-C14-HSL, promote plant growth, treatments with C6-HSL, oxo-C10-HSL, or oxo-C14-HSL resulted in different transcriptional profiles in Arabidopsis. Understanding the priming induced by bacterial QS molecules augments our knowledge of plant reactions to bacteria and suggests strategies for using beneficial bacteria in plant protection.

Measurement Properties of Mean Length of Utterance in School-Age Children.

PURPOSE: Mean length of utterance (MLU) is one of the most widely reported measures of syntactic development in the developmental literature, but its responsiveness in young school-age children's language has been questioned, and it has been shown to correlate with nonsyntactic measures. This study tested the extent to which MLU shows measurement properties of responsiveness and construct validity when applied to language elicited from elementary school children. METHOD: Thirty-two typically developing children in two age groups (5 and 8 years) provided four short language samples each. Language samples were elicited in a question-answer context and a narrative context. MLU was calculated with both morpheme and word counts. Other established measures of syntactic complexity (clausal density [CD], developmental level [D-Level], mean length of clause [MLC]) and lexical diversity (lexical density, moving-average type-token ratio, number of different words) were also calculated. RESULTS: Linear mixed-effects analyses revealed that MLU varied systematically with discourse context and children's age group. The syntactic measures, CD and MLC, were found to vary systematically with MLU. None of the lexical diversity measures varied systematically with MLU. CONCLUSION: Results suggest that MLU is a responsive and valid measure of children's syntactic development across age and discourse context during the early school-age years.

INVESTIGATING METRICAL CONTEXT EFFECTS ON ANTICIPATORY COARTICULATION IN CONNECTED SPEECH DEVELOPMENT.

If rhythm acquisition is influenced by the development of articulatory timing, then metrical structure might be expected to condition this timing. This study tested this hypothesis by investigating anticipatory effects of an upcoming noun on the production of a preceding determiner, under the assumption that anticipatory coarticulation indexes chunking. Simple S-V-O sentences were elicited from 5-year-olds, 8-year-olds, and adults. The V was either monosyllabic packed or disyllabic patted. The O was a determiner phrase where nouns varied either in onset place-of-articulation (POA; tack vs. cat) or in their rhymes (tack vs. toot). Acoustic analyses of determiner schwa F1 and F2 showed no effect of verb on schwa coarticulation. Given other results, including an interaction between age group and POA, the findings suggest that the acquisition of articulatory timing is independent of metrical structure, even if this timing is related to speech rhythm acquisition.

Bang for Your Buck: A Single-Case Experimental Design Study of Practice Amount and Distribution in Treatment for Childhood Apraxia of Speech.

Purpose The aim of this study was to examine 2 aspects of treatment intensity in treatment for childhood apraxia of speech (CAS): practice amount and practice distribution. Method Using an alternating-treatments single-subject design with multiple baselines, we compared high versus low amount of practice, and massed versus distributed practice, in 6 children with CAS. Conditions were manipulated in the context of integral stimulation treatment. Changes in perceptual accuracy, scored by blinded analysts, were quantified with effect sizes. Results Four children showed an advantage for high amount of practice, 1 showed an opposite effect, and 1 showed no condition difference. For distribution, 4 children showed a clear advantage for massed over distributed practice post treatment; 1 showed an opposite pattern, and 1 showed no clear difference. Follow-up revealed a similar pattern. All children demonstrated treatment effects (larger gains for treated than untreated items). Conclusions High practice amount and massed practice were associated with more robust speech motor learning in most children with CAS, compared to low amount and distributed practice, respectively. Variation in effects across children warrants further research to determine factors that predict optimal treatment conditions. Finally, this study adds to the evidence base supporting the efficacy of integral stimulation treatment for CAS. Supplemental Material https://doi.org/10.23641/asha.9630599.

English speech sound development in preschool-aged children from bilingual English-Spanish environments.

PURPOSE: English speech acquisition by typically developing 3- to 4-year-old children with monolingual English was compared to English speech acquisition by typically developing 3- to 4-year-old children with bilingual English-Spanish backgrounds. We predicted that exposure to Spanish would not affect the English phonetic inventory but would increase error frequency and type in bilingual children. METHOD: Single-word speech samples were collected from 33 children. Phonetically transcribed samples for the 3 groups (monolingual English children, English-Spanish bilingual children who were predominantly exposed to English, and English-Spanish bilingual children with relatively equal exposure to English and Spanish) were compared at 2 time points and for change over time for phonetic inventory, phoneme accuracy, and error pattern frequencies. RESULTS: Children demonstrated similar phonetic inventories. Some bilingual children produced Spanish phonemes in their English and produced few consonant cluster sequences. Bilingual children with relatively equal exposure to English and Spanish averaged more errors than did bilingual children who were predominantly exposed to English. Both bilingual groups showed higher error rates than English-only children overall, particularly for syllable-level error patterns. All language groups decreased in some error patterns, although the ones that decreased were not always the same across language groups. Some group differences of error patterns and accuracy were significant. Vowel error rates did not differ by language group. CONCLUSION: Exposure to English and Spanish may result in a higher English error rate in typically developing bilinguals, including the application of Spanish phonological properties to English. Slightly higher error rates are likely typical for bilingual preschool-aged children. Change over time at these time points for all 3 groups was similar, suggesting that all will reach an adult-like system in English with exposure and practice.

Cross-linguistic generalization in the treatment of two sequential Spanish-English bilingual children with speech sound disorders.

PURPOSE: The effect of bilingual service delivery on treatment of speech sound disorders (SSDs) in bilingual children is largely unknown. Bilingual children with SSDs are typically provided intervention in only one language, although research suggests dual-language instruction for language disorders is best practice for bilinguals. This study examined cross-linguistic generalization of bilingual intervention in treatment of two 5-year-old sequential bilingual boys with SSDs (one with Childhood Apraxia of Speech), hypothesizing that selecting and treating targets in both languages would result in significant overall change in their English and Spanish speech systems. METHOD: A multiple baseline across behaviours design was used to measure treatment effectiveness for two targets per child. Children received treatment 2-3 times per week for 8 weeks and in Spanish for at least 2 of every 3 days. Ongoing treatment performance was measured in probes in both languages; overall speech skills were compared pre- and post-treatment. RESULT: Both children's speech improved in both languages with similar magnitude; there was improvement in some non-treated errors. CONCLUSION: Treating both languages had an overall positive effect on these bilingual children's speech. Future bilingual intervention research should explore alternating treatments designs, efficiency of monolingual vs bilingual treatment, different language and bilingual backgrounds, and between-group comparisons.

N-acyl-homoserine lactones-producing bacteria protect plants against plant and human pathogens.

The implementation of beneficial microorganisms for plant protection has a long history. Many rhizobia bacteria are able to influence the immune system of host plants by inducing resistance towards pathogenic microorganisms. In this report, we present a translational approach in which we demonstrate the resistance-inducing effect of Ensifer meliloti (Sinorhizobium meliloti) on crop plants that have a significant impact on the worldwide economy and on human nutrition. Ensifer meliloti is usually associated with root nodulation in legumes and nitrogen fixation. Here, we suggest that the ability of S. meliloti to induce resistance depends on the production of the quorum-sensing molecule, oxo-C14-HSL. The capacity to enhanced resistance provides a possibility to the use these beneficial bacteria in agriculture. Using the Arabidopsis-Salmonella model, we also demonstrate that the application of N-acyl-homoserine lactones-producing bacteria could be a successful strategy to prevent plant-originated infections with human pathogens.

Motor-based intervention protocols in treatment of childhood apraxia of speech (CAS).

This paper reviews current trends in treatment for childhood apraxia of speech (CAS), with a particular emphasis on motor-based intervention protocols. The paper first briefly discusses how CAS fits into the typology of speech sound disorders, followed by a discussion of the potential relevance of principles derived from the motor learning literature for CAS treatment. Next, different motor-based treatment protocols are reviewed, along with their evidence base. The paper concludes with a summary and discussion of future research needs.

The Salmonella effector protein SpvC, a phosphothreonine lyase is functional in plant cells.

Salmonella is one of the most prominent causes of food poisoning and growing evidence indicates that contaminated fruits and vegetables are an increasing concern for human health. Successful infection demands the suppression of the host immune system, which is often achieved via injection of bacterial effector proteins into host cells. In this report we present the function of Salmonella effector protein in plant cell, supporting the new concept of trans-kingdom competence of this bacterium. We screened a range of Salmonella Typhimurium effector proteins for interference with plant immunity. Among these, the phosphothreonine lyase SpvC attenuated the induction of immunity-related genes when present in plant cells. Using in vitro and in vivo systems we show that this effector protein interacts with and dephosphorylates activated Arabidopsis Mitogen-activated Protein Kinase 6 (MPK6), thereby inhibiting defense signaling. Moreover, the requirement of Salmonella SpvC was shown by the decreased proliferation of the ΔspvC mutant in Arabidopsis plants. These results suggest that some Salmonella effector proteins could have a conserved function during proliferation in different hosts. The fact that Salmonella and other Enterobacteriaceae use plants as hosts strongly suggests that plants represent a much larger reservoir for animal pathogens than so far estimated.

The importance of production frequency in therapy for childhood apraxia of speech.

PURPOSE: This study explores the importance of production frequency during speech therapy to determine whether more practice of speech targets leads to increased performance within a treatment session, as well as to motor learning, in the form of generalization to untrained words. METHOD: Two children with childhood apraxia of speech were treated with an alternating treatment AB design, with production frequency differing in the 2 treatments. The higher production frequency treatment required 100+ productions in 15 min, while the moderate-frequency treatment required 30-40 productions in the same time period. One child was treated 3 times weekly for 11 weeks; the other child was treated twice weekly for 5 weeks. At the conclusion of each treatment phase, 5 min of probes were administered to determine whether generalization had occurred. Maintenance data to measure performance and learning were collected after a break from treatment. RESULTS: Both children showed improvement on all targets; however, the targets with the higher production frequency treatment were acquired faster, evidenced by better in-session performance and greater generalization to untrained probes. CONCLUSIONS: Both treatment designs were effective, though frequent and intense practice of speech resulted in more rapid response to treatment in 2 children whose primary communication difficulty was childhood apraxia of speech.

English speech acquisition in 3- to 5-year-old children learning Russian and English.

PURPOSE: English speech acquisition in Russian-English (RE) bilingual children was investigated, exploring the effects of Russian phonetic and phonological properties on English single-word productions. Russian has more complex consonants and clusters and a smaller vowel inventory than English. METHOD: One hundred thirty-seven single-word samples were phonetically transcribed from 14 RE and 28 English-only (E) children, ages 3;3 (years;months) to 5;7. Language and age differences were compared descriptively for phonetic inventories. Multivariate analyses compared phoneme accuracy and error rates between the two language groups. RESULTS: RE children produced Russian-influenced phones in English, including palatalized consonants and trills, and demonstrated significantly higher rates of trill substitution, final devoicing, and vowel errors than E children, suggesting Russian language effects on English. RE and E children did not differ in their overall production complexity, with similar final consonant deletion and cluster reduction error rates, similar phonetic inventories by age, and similar levels of phonetic complexity. Both older language groups were more accurate than the younger language groups. CONCLUSIONS: We observed effects of Russian on English speech acquisition; however, there were similarities between the RE and E children that have not been reported in previous studies of speech acquisition in bilingual children. These findings underscore the importance of knowing the phonological properties of both languages of a bilingual child in assessment.

The development of distinct speaking styles in preschool children.

PURPOSE: To examine when and how socially conditioned distinct speaking styles emerge in typically developing preschool children's speech. METHOD: Thirty preschool children, ages 3, 4, and 5 years old, produced target monosyllabic words with monophthongal vowels in different social-functional contexts designed to elicit clear and casual speaking styles. Thirty adult listeners were used to assess whether and at what age style differences were perceptible. Children's speech was acoustically analyzed to evaluate how style-dependent differences were produced. RESULTS: The ratings indicated that listeners could not discern style differences in 3-year-olds' speech but could hear distinct styles in 4-year-olds' and especially in 5-year-olds' speech. The acoustic measurements were consistent with these results: Style-dependent differences in 4- and 5-year-olds' words included shorter vowel durations and lower fundamental frequency in clear compared with casual speech words. Five-year-olds' clear speech words also had more final stop releases and initial sibilants with higher spectral energy than did their casual speech words. Formant frequency measures showed no style-dependent differences in vowel production at any age nor any differences in initial stop voice onset times. CONCLUSION: Overall, the findings suggest that distinct styles develop slowly and that early style-dependent differences in children's speech are unlike those observed in adult clear and casual speech. Children may not develop adultlike styles until they have acquired expert articulatory control and the ability to highlight the internal structure of an articulatory plan for a listener.

The acquisition of two phonetic cues to word boundaries.

The study evaluated whether durational and allophonic cues to word boundaries are intrinsic to syllable production, and so acquired with syllable structure, or whether they are suprasyllabic, and so acquired in phrasal contexts. Twenty preschool children (aged 3;6 and 4;6) produced: (1) single words with simple and complex onsets (e.g. nail vs. snail); and (2) two-word phrases with intervocalic consonant sequences and varying boundary locations (e.g. this nail vs. bitty snail). Comparisons between child and adult control productions showed that the durational juncture cue was emergent in the four-year-olds' productions of two-word phrases, but absent elsewhere. In contrast, the allophonic cue was evident even in the three-year-olds' productions of single words. Perceptual judgments showed that age- and type-dependent acoustic differences translated into differences in listener behavior. The differential acquisition of the two juncture cues is discussed with reference to the acquisition of articulatory timing control.

The endophytic fungus Piriformospora indica reprograms barley to salt-stress tolerance, disease resistance, and higher yield.

Disease resistance strategies are powerful approaches to sustainable agriculture because they reduce chemical input into the environment. Recently, Piriformospora indica, a plant-root-colonizing basidiomycete fungus, has been discovered in the Indian Thar desert and was shown to provide strong growth-promoting activity during its symbiosis with a broad spectrum of plants. Here, we report on the potential of P. indica to induce resistance to fungal diseases and tolerance to salt stress in the monocotyledonous plant barley. The beneficial effect on the defense status is detected in distal leaves, demonstrating a systemic induction of resistance by a root-endophytic fungus. The systemically altered "defense readiness" is associated with an elevated antioxidative capacity due to an activation of the glutathione-ascorbate cycle and results in an overall increase in grain yield. Because P. indica can be easily propagated in the absence of a host plant, we conclude that the fungus could be exploited to increase disease resistance and yield in crop plants.